Professional Documents
Culture Documents
in
Microbiology
- Module: Prokaryotes -
Contents
1. INTRODUCTION
1.1 Rules of operation of the laboratory of practical classes
1.2 Asepsis procedures
1.3 Preparation of media and materials
Bibliography
1. Cappuccino, J.G. & Welsh, C. 2018. Microbiology: a Laboratory Manual. 11th ed.
Harlow: Pearson Education.
2. Harley, J.P. & Prescott, L.M. 2002. Laboratory Exercises in Microbiology. 5th ed.
New York: McGraw-Hill Company
3. Meynell, G.G. & Meynell, E. 1970. Theory and Practice in Experimental Bacteriology.
2nd ed. Cambridge: Cambridge University Press.
4. Murray, P.R., Baron, E.J., Pfaller, M.A., Tenover, F.C. & Yolken, R.H. , Eds. 1995.
Manual of Clinical Microbiology. 6th ed. Washington: ASM Press.
5. Reddy, C.A., Beveridge, T.J., Breznak, J.A., Marzluf, G.A., Schmidt, T.M.,Snyder,
L.R. (Eds.). 2007. Methods for General and Molecular Microbiology. 3rd ed.
Washington: ASM.
• Cap 1: Light microscopy (pp. 3 a 18)
• Cap 2: Sampling and staining for light microscopy (pp. 19 a 33)
• Cap 9: Growth measurement (pp. 172 a 199)
• Cap 15 : Phenotypic characterization and the principles of comparative
systematics (pp. 341, 342, 350-351, 352, 367, 381)
6. Versalovic, J. (Editor in Chief). 2011. Manual of Clinical Microbiology. 10th ed.
Washington: ASM Press.
Master in Microbiology
PW1
LABORATORY TECHNIQUES IN BACTERIOLOGY
See
1. PRACTICAL PROGRAMME
SECTION 2. LABORATORY TECHNIQUES IN BACTERIOLOGY [PW1]
Isolamento
de
microrganismos
|
Tipos
morfo-‐fisiológicos
Tipo$I$ Tipo$II$ Tipo$III$ Tipo$IV$ Tipo$V$ Tipo$VI$ Tipo$VII$ Tipo$VIII$ Tipo$IX$ Tipo$X$ Tipo$XI$
Espécie/estirpe
Diâmetro
Pigmentação
Consistência
Morfologia Opacidade
Colónias Textura
Forma
Elevação
Margem
Forma
Morfologia
Agrupamento
Celular
Dimensões
Parede Gram
Celular KOH
Forma
Posição
Endósporos
Dilatação
Popping
Negativa
Colorações
Negro do Sudão
Mobilidade
Caracteres
Catalase
Fisiológicos
Oxidase
Master in Microbiology
PW2
ISOLATION AND IDENTIFICATION OF Bacillus spp. STRAINS
Master in Microbiology
1. Isolation
1.1 For each supplied sample prepare, under aseptic conditions, a 10% w/v suspension in
10 ml of saline solution (0.9% NaCl) or peptonated water (0.1% peptone).
1.2 Homogenise the suspension by vigorous vortexing and proceed with the pasteurisation
process: place the suspension in a water bath at 80°C for 10-15 min, then cool on ice.
1.3 After stirring the pasteurized suspension by vortexing, plate an aliquot on an ioslation
medium plate: TSA medium (triptone 17 g/L; peptone 3 g/L; glucose 2,5 g/L; dipotassium
phosphate 2,5 g/L; sodium chloride 5 g/L; agar 15 g/L) or NA medium (tryptone 10 g/L,
meat extract 5 g/L; sodium chloride 5 g/L; agar 15 g/L).
To obtain isolated colonies, dilute previously (10-2) and plate 0.1 ml or, alternatively,
inoculate a loop of the original suspension.
1.5 When verifying the appearance of colonies, microscopic observation should be carried
out to detect the presence of gram-positive bacilli, catalase positive and endospore
producers.
Select two colonies which, taking into account the characteristics described for the genus
Bacillus, may belong to different species.
1.6 Isolate each of the two selected strains in pure culture by successive passages
(at least 3) by non-selective media (nutrient agar, NA). Between each passage, incubate
the plates at isolation temperature.
1.7 At the end of the isolation, confirm that the bacilli are gram positive, catalase positive
and endospore producers. Refer to the strains with the code BACxA and BACxB, where x is
the student group number.
2. Identification
2.1. For the strain of your group that is selected in the class, carry out its identification at
species level, using the miniaturized galleries API 50CH and the suspension medium API
50CHB, according to the manufacturer's instructions.
2.2 In parallel, use the test results of the API50CHB galleries and carry out the probabilistic
identification according to the teacher's instructions.
2.3 If it is impossible to achieve an identification, check that the final isolation plate does
not have a mixed culture (failure of the purification process).
Master in Microbiology
1. Isolation
1.1 For each supplied sample prepare, under aseptic conditions, a 10% w/v suspension in
10 ml of saline solution (0.9% NaCl) or peptonated water (0.1% peptone).
1.2 Homogenise the suspension by vigorous vortexing and proceed with the pasteurisation
process: place the suspension in a water bath at 80°C for 10-15 min, then cool on ice.
1.3 After stirring the pasteurized suspension by vortexing, plate an aliquot on an ioslation
medium plate: TSA medium (triptone 17 g/L; peptone 3 g/L; glucose 2,5 g/L; dipotassium
phosphate 2,5 g/L; sodium chloride 5 g/L; agar 15 g/L) or NA medium (tryptone 10 g/L,
meat extract 5 g/L; sodium chloride 5 g/L; agar 15 g/L).
To obtain isolated colonies, dilute previously (10-2) and plate 0.1 ml or, alternatively,
inoculate a loop of the original suspension.
1.5 When verifying the appearance of colonies, microscopic observation should be carried
out to detect the presence of gram-positive bacilli, catalase positive and endospore
producers.
Select two colonies which, taking into account the characteristics described for the genus
Bacillus, may belong to different species.
1.6 Isolate each of the two selected strains in pure culture by successive passages
(at least 3) by non-selective media (nutrient agar, NA). Between each passage, incubate
the plates at isolation temperature.
1.7 At the end of the isolation, confirm that the bacilli are gram positive, catalase positive
and endospore producers. Refer to the strains with the code BACxA and BACxB, where x is
the student group number.
2. Identification
2.1. For the strain of your group that is selected in the class, carry out its identification at
species level, using the miniaturized galleries API 50CH and the suspension medium API
50CHB, according to the manufacturer's instructions.
2.2 In parallel, use the test results of the API50CHB galleries and carry out the probabilistic
identification according to the teacher's instructions.
2.3 If it is impossible to achieve an identification, check that the final isolation plate does
not have a mixed culture (failure of the purification process).
Tabela de Diagnóstico do género Bacillus
Espécies
Ø > 1 µm
Mobilidade
E. redondo
E. elipsoide
E. cilindrico
E. central
E. sub-terminal
E. dilatado
Vacuolos
D-glucose
Glc. Anaerobiose
L-arabinose
D-arabinose
D-xilose
D-manitol
Galactose
Ribose
Glicerol
D-manose
Ramnose
Inositol
D-rafinose
Lactose
Citrato
Amido
Gelatina
Caseína
Esculina
Urease
Catalase
NO 3 →NO 2
Teste VP
pH < 6 (VP)
pH > 7 (VP)
Cresc. pH 5,7
Cresc. NACl 7%
B. cereus 9 9 1 9 1 5 9 1 9 9 9 1 1 1 1 1 9 9 1 1 1 1 1 9 9 9 9 9 5 9 9 9 9 1 9 5
B. mycoides 9 1 1 9 1 9 9 1 9 9 9 1 1 1 5 5 5 9 1 1 1 1 1 5 9 9 9 9 5 9 5 9 9 1 9 5
B. thuringiensis 9 9 1 9 1 1 9 1 9 9 9 1 1 1 1 1 9 9 5 1 1 1 1 9 9 9 9 9 1 9 9 9 9 1 9 9
B. anthracis 9 1 1 9 5 1 9 1 9 9 9 1 1 1 1 1 9 1 1 1 1 1 1 1 9 5 9 9 1 9 9 9 9 1 9 9
B. firmus 1 9 1 9 1 5 9 5 1 9 1 1 1 1 9 1 5 9 1 1 1 1 1 5 9 9 9 9 1 9 5 5 1 1 1 9
B. lentus 1 9 1 9 1 5 9 5 1 9 1 9 1 9 9 5 9 5 9 5 1 9 5 1 9 5 5 9 5 9 1 5 1 5 1 5
B. laterosporus 1 9 1 9 1 9 9 9 1 9 9 1 1 1 9 1 9 9 9 1 1 1 1 5 5 9 9 9 1 9 1 5 5 1 1 1
B. alvei 1 9 1 9 1 5 9 9 1 9 9 1 1 1 1 5 9 9 5 1 5 5 1 1 9 9 9 9 5 9 1 9 9 1 1 1
B. brevis 1 9 1 9 1 9 9 9 1 5 1 1 1 1 5 1 1 5 1 1 1 1 1 5 1 5 9 1 1 9 9 5 1 9 5 1
B. sphaericus 1 9 9 5 1 1 5 9 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 5 1 5 5 1 5 9 1 1 1 9 5 5
B. subtilis 1 9 1 9 1 5 9 1 1 9 1 9 1 9 9 1 9 9 9 1 9 9 5 9 9 9 9 9 1 9 9 9 5 1 9 9
B. licheniformis 1 9 1 9 5 5 9 1 1 9 9 9 1 9 9 9 9 9 9 9 5 9 9 9 9 9 9 9 5 9 9 9 9 1 9 9
B. pumilus 1 9 1 5 9 5 9 1 1 9 1 9 1 9 9 9 9 9 9 1 5 9 5 9 1 9 9 9 1 9 1 9 9 1 9 9
B. megaterium 9 9 5 9 1 5 9 1 1 9 1 5 1 5 5 9 9 9 1 1 9 9 9 9 9 9 9 9 1 9 1 5 5 1 5 5
B. circularis 1 9 1 9 5 5 9 9 1 9 5 9 5 9 9 9 9 9 9 5 5 9 9 1 9 5 5 9 1 9 5 5 9 1 5 5
B. macerans 1 9 1 9 1 1 9 9 1 9 9 9 9 9 9 9 9 9 9 9 5 9 9 1 9 5 1 9 1 9 5 5 9 1 9 1
B. polymyxa 1 9 1 9 1 5 9 9 1 9 9 9 1 9 9 9 9 9 9 5 1 9 9 1 9 9 9 9 1 9 1 9 5 1 9 1
B. pantothenticus 1 9 5 5 1 1 5 9 1 9 9 1 9 1 1 9 9 9 9 9 5 1 5 5 9 9 5 9 1 9 5 5 9 1 1 9
B. coagulans 1 9 1 5 1 1 9 9 1 9 9 5 1 5 5 9 9 9 9 5 5 9 9 1 9 5 5 9 1 9 1 9 9 1 9 1
B. stearothermophilus 1 9 1 9 1 1 9 9 1 9 1 5 1 5 5 5 5 9 9 1 5 5 5 1 5 9 5 5 1 5 5 5 9 1 1 1
REF 50 300 07945F - GB - 2002/11
®
50 CH IVD
Carbohydrates
bioMérieux® sa English - 1
api® 50 CH 07945F - GB - 2002/11
REAGENTS AND MATERIAL REQUIRED BUT NOT SPECIMENS (COLLECTION AND PREPARATION)
PROVIDED API 50 CH is not for use directly with clinical or other
Reagents : specimens.
- Inoculation medium : The microorganisms to be identified must first be isolated
API 50 CHL Medium (Ref. 50 410) on a suitable culture medium according to standard
API 50 CHB/E Medium (Ref. 50 430) microbiological techniques.
(+ products mentioned in the package inserts
of these media) INSTRUCTIONS FOR USE
or any other suitable medium Depending on which medium is used, API 50 CHL
- Mineral oil (Ref. 70 100) Medium or API 50 CHB/E Medium, carefully read the
- McFarland Standard (Ref. 70 900) or corresponding package insert.
DENSIMAT (Ref. 99 234) or ATB® Densitometer
- Identification software (consult bioMérieux) Preparation of the strips
Each strip is made up of 5 smaller strips each containing
Material :
10 numbered tubes.
- Pipettes or PSIpettes
• Prepare an incubation box (tray and lid).
- Ampule rack
• Record the reference of the strain on the elongated flap
- Ampule protector
of the tray. (Do not record the reference on the lid as it
- Swabs
may be misplaced during the procedure.)
- General microbiology laboratory equipment
• Distribute about 10 ml of distilled water or de-
mineralized water [or any water without additives or
WARNINGS AND PRECAUTIONS chemicals which may release gases (e.g. Cl2, CO2, etc.)]
• For in vitro diagnostic use and microbiological into the honeycombed wells of the tray to create a
control. humid atmosphere.
• For professional use only. • Remove the 2 strips (0-19 and 20-39) from their
• This kit contains products of animal origin. Certified packaging, separate into 4 smaller strips (0-9, 10-19,
knowledge of the origin and/or sanitary state of the 20-29 and 30-39) and place all 4 in the incubation tray.
animals does not totally guarantee the absence of • Take the remaining smaller strip (40-49) out of the
transmissible pathogenic agents. It is therefore packaging and place it next to the others in the
recommended that these products be treated as incubation tray to complete the strip.
potentially infectious, and handled observing the usual
safety precautions (do not ingest or inhale). Preparation of the inoculum
• All specimens, microbial cultures and inoculated • Culture the microorganism on a medium adapted to its
products should be considered infectious and handled growth.
appropriately. Aseptic technique and usual precautions • Check the purity of the strain.
for handling the bacterial group studied should be • Harvest all the bacteria from a solid medium using a
observed throughout this procedure. Refer to "NCCLS swab or from a liquid medium using centrifugation.
M29-A, Protection of Laboratory Workers from • Prepare the inoculum in the appropriate medium (see
Instrument Biohazards and Infectious Disease the API 50 CHL Medium and API 50 CHB/E Medium
Transmitted by Blood, Body Fluids, and Tissue; package inserts).
Approved Guideline - December 1997". For additional This suspension must be used immediately after
handling precautions, refer to "Biosafety in preparation.
Microbiological and Biomedical Laboratories, HHS
Inoculation of the strips
Publication No. (CDC) 93-8395, 3rd Edition (May
1993)", or to the regulations currently in use in each Distribute the bacterial suspension using a sterile pipette
country. into the 50 tubes as follows :
• Do not use reagents past the expiration date. • Tilt the incubation box slightly forwards.
• Before use, check that the packaging of the various • Avoid the formation of bubbles by placing the tip of the
components is intact. pipette against the side of the cupule.
• Do not use strips which have been damaged : cupules • When only the tube is to be inoculated, do not exceed
deformed, desiccant sachet open, ... the top of the tube so as to maintain anaerobic
• The performance data were obtained using the conditions.
procedure indicated in this package insert. Any change • When the tube and the cupule are to be completely
or modification in the procedure may affect the results. filled, avoid the formation of a concave or convex
• Interpretation of the test results should be made taking meniscus.
into consideration the patient history, the source of the • Incubate the strips at the optimum temperature for
specimen, colonial and microscopic morphology of the growth of the group of microorganisms being tested :
strain and, if necessary, the results of any other tests 30°C, 37°C or 55°C.
performed, particularly the antimicrobial susceptibility
patterns.
STORAGE CONDITIONS
The strips should be stored at 2-8°C until the expiration
date indicated on the packaging.
bioMérieux® sa English - 2
api® 50 CH 07945F - GB - 2002/11
READING AND INTERPRETATION The results form a biochemical profile which, when
Reading the strips entered in the identification software, provides the
identification of Lactobacillus and related genera or
The strips are read after the stipulated incubation times
Bacillus and related genera, Enterobacteriaceae and
(e.g., 24 hrs., 48 hrs.), depending on the microorganism
Vibrionaceae.
and the type of reaction studied.
NOTE : The results may be used for other purposes :
Interpretation • Epidemiological grouping into types of the
Interpret each test (positive (+), negative (-), doubtful (?)) microorganism.
and record the results on the result sheet. • Taxonomical analysis of a group of microorganisms.
• Classification of an unknown bacterial population into
homogeneous groups.
QUALITY CONTROL
The strips are systematically controlled at various stages of their manufacture. For those users who wish to perform their
own quality control tests with the strip, the following strains may be used :
For Lactobacillus : Lactobacillus paracasei ssp paracasei NFCB 206 or ATCC BAA-52 (with API 50 CHL Medium)
0 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49
24 - - - - - + - - - - + + + + - - V - + V - + + - V + V V + - - + + + + - - - - - + - + - - - - V - -
48 - - - - - + - - - - + + + + - - + - + + - + + V + + + + + - - + + + + - - - - V + - + - - - - + - -
For Bacillus : Bacillus polymyxa (*) ATCC 43865 (with API 50 CHB/E Medium)
0 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49
24 - + - - + + + - - V + + + + - V - - + - - + - + + + + + + + + + + - + + + + - + + - - - - - - - - -
48 - + - - + + + - - + + + + + - V - - + - - + - + + + + + + + + + + V + + + + - + + - - - - - - - - -
(*) Bacillus polymyxa identified as Paenibacillus polymyxa with API 50 CH and API 50 CHB/E Medium.
Results obtained after incubation at 30°C.
For Enterobacteriaceae : Klebsiella pneumoniae ssp pneumoniae ATCC 35657 (with API 50 CHB/E Medium)
0 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49
24 - + - - + + + - + - + + + + - + - + + + - + + - + + + + + + + + + - - + + - - + - - - - + + - + + -
48 - + - V + + + - + - + + + + - + - + + + - + + V + + + + + + + + + - - + + - - + V - - - + + - + + -
ATCC : American Type Culture Collection, 10801 University Boulevard, Manassas, VA 20110-2209, USA.
NCFB : National Collection for Food Bacteria (=NCDO), Institute of Food Research, Reading Laboratory, Earley Gate, Reading
RG6 6BZ, ENGLAND
It is the responsibility of the user to perform Quality Control in accordance with any local applicable regulations.
LITERATURE REFERENCES p. I
INDEX OF SYMBOLS p. II
®
50 CH IVD
Carbohidratos
bioMérieux® sa Português - 1
api® 50 CH 07945F - PT - 2002/11
CONDIÇÕES DE ARMAZENAMENTO
As galerias conservam-se a 2º-8ºC até à data de validade
indicada na embalagem.
bioMérieux® sa Português - 2
api® 50 CH 07945F - PT - 2002/11
Para Bacillus : com a estirpe/cepa Bacillus polymyxa (*) ATCC 43865 (com API 50 CHB/E Medium)
0 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49
24 - + - - + + + - - V + + + + - V - - + - - + - + + + + + + + + + + - ++ + + - + + - - - - - - - - -
48 - + - - + + + - - + + + + + - V - - + - - + - + + + + + + + + + + V + + + + - + + - - - - - - - - -
(*) Bacillus polymyxa identificado a Paenibacillus polymyxa em API 50 CH e API 50 CHB/E Medium.
Resultados obtidos após incubação a 30°C.
Para Enterobacteriaceae : com a estirpe/cepa Klebsiella pneumoniae ssp pneumoniae ATCC 35657 (com API 50
CHB/E Medium)
0 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49
24 - + - - + + + - + - + + + + - + - + + + - + + - + + + + + + + + + - - + + - - + - - - - + + - + + -
48 - + - V + + + - + - + + + + - + - + + + - + + V + + + + + + + + + - - + + - - + V - - - + + - + + -
ATCC : American Type Culture Collection, 10801 University Boulevard, Manassas, VA 20110-2209, USA.
NCFB : National Collection for Food Bacteria (=NCDO), Institute of Food Research, Reading Laboratory, Earley Gate, Reading
RG6 6BZ, ENGLAND
É da responsabilidade do utilizador assegurar que o controlo de qualidade é efectuado em conformidade com a
legislação local em vigor.
LIMITES DO TESTE COMPORTAMENTO FUNCIONAL
• Qualquer identificação de espécies não enumeradas Consultar o comportamento funcional dos meios
nas bases de dados API 50 CHL e API 50 CHB/E está associados a esta galeria.
sob a responsabilidade do utilizador.
ELIMINAÇÃO DE RESÍDUOS
• Devem apenas ser utilizadas culturas puras contendo
um único tipo de microrganismo. É da responsabilidade de cada laboratório gerir os
resíduos e os efluentes que este produz consoante a sua
RESULTADOS ESPERADOS natureza e o seu perigo, e assegurar (ou fazer assegurar)
Consultar o Quadro de Identificação no final deste folheto o tratamento e a eliminação em conformidade com as
informativo para saber os resultados esperados para as regulamentações aplicáveis.
diferentes reacções bioquímicas.
BIBLIOGRAFIA p. I
QUADRO DE SÍMBOLOS p. II
Brasil: Distribuído por biolab-Mérieux, S.A. - Estrada do Mapuá, 491 - Jacarepaguá - R.J. - CEP 22710-261
CNPJ: 33.040.635/0001-71
Atendimento ao Consumidor Tel.: 0800-264848
Prazo de Validade, N° de Lote, N° de Registro de Ministério da Saúde e Responsável Técnico:
VIDE EMBALAGEM
®
50 CHB/E Medium IVD
bioMérieux SA English - 1
api® 50 CHB/E Medium 07964G - en - 2011/07
SPECIMENS (COLLECTION AND PREPARATION) If the DENSIMAT or ATB Densitometer is not used :
API 50 CHB/E Medium is not for use directly with clinical 1) Suspension for inoculation of the API 20 E strip :
or other specimens. - Open a tube containing 1 ml of sterile saline.
The microorganisms to be identified must first be isolated - Pick up all the bacteria from the culture using a
on a suitable culture medium according to standard swab.
microbiological techniques. - Prepare a heavy suspension (S) in the tube.
- Open an ampule of API NaCl 0.85 % Medium
INSTRUCTIONS FOR USE (5 ml) as indicated in the paragraph "Warnings and
Precautions".
For Bacillus - Prepare a suspension with a turbidity equivalent to
2 McFarland by transferring a certain number of
Selection of the colonies drops of suspension S into the ampule : record this
Check the purity of the strain. number of drops (n).
Check that it belongs to the Bacillus genus : aerobic, 2) Suspension for inoculation of the API 50 CH strip :
spore-forming rod, usually Gram-positive. - Open an ampule of API 50 CHB/E Medium as
Culture it on a nutrient agar plate. indicated in the paragraph "Warnings and
- If the optimum growth temperature of the micro- Precautions".
organism is unknown, incubate several plates at - Inoculate the ampule of API 50 CHB/E Medium by
different temperatures. transferring twice the number of drops of
- For slow growing strains, use two plates so as to have suspension (i.e. 2n) into the ampule.
enough bacteria : Homogenize.
- mesophiles grow at temperatures between 25°C and
45°C during 16-18 hours ; NOTE : If the API 20 E strip is be used in association with
- psychrophiles grow at 20°C during 18-48 hours ; the API 50 CH strip, follow the instructions in the API 20 E
- thermophiles grow at 55°C during 12-16 hours. package insert.
Growth of Bacillus lentus is encouraged by the addition Inoculation of the strips
of 1 g of urea/litre into the nutrient agar before
(see the API 50 CH and API 20 E package inserts)
sterilization.
Preparation of the strips Fill the tubes (not the cupules) with the inoculated
API 50 CHB/E Medium.
See the package inserts for API 50 CH and API 20 E (use NOTE : The addition of mineral oil is optional ; it is
optional). not however recommended for strict aerobic
Preparation of the inoculum bacteria.
Inoculate the first 12 tests only of the API 20 E strip, as
The solutions must be used immediately after preparation.
the last 8 tests are duplicated on the API 50 CH strip,
If the DENSIMAT or ATB TM Densitometer is used : and inoculate the GLU test to reveal the NIT reaction.
1) Suspension for inoculation of the API 50 CH strip :
Incubation of the strips
- Open an ampule of API 50 CHB/E Medium as
indicated in the paragraph "Warnings and Incubate :
Precautions". - thermophilic species at 55°C ± 2°C for 3 - 3 ½ hours,
- Pick up several identical colonies. 6 - 6 ½ hours and 24 hours (± 2 hours), slightly tilting
- Prepare a suspension with a turbidity equivalent to the API 50 CH strip, base of the tubes uppermost, in
2 McFarland in the ampule of API 50 CHB/E order to trap any gas produced,
Medium. - other species at 29°C ± 2°C for 24 hours (± 2 hours)
and 48 hours (± 6 hours).
2) Suspension for inoculation of the API 20 E strip :
NOTE : For the API 20 E strip, the same incubation
- Open an ampule of API NaCl 0.85 % Medium
conditions must be observed.
(5 ml) as indicated in the paragraph "Warnings and
Precautions". Reading the strips
- Pick up several identical colonies. (see the API 50 CH and API 20 E package inserts)
- Prepare a suspension with a turbidity equivalent to
2 McFarland. Read the results :
- for thermophilic species after 3 - 3 ½ hours, 6 - 6 ½
hours and 24 hours (± 2 hours) of incubation,
- for other species after 24 hours (± 2 hours) and
48 hours (± 6 hours) of incubation.
For the API 50 CH strip :
- A positive test corresponds to acidification revealed by
the phenol red indicator contained in the medium
changing to YELLOW.
- For the esculin test (tube no. 25), a change in color
from red to BLACK is observed.
NOTE : If a positive test becomes negative at the
second reading, only the positive result should be taken
into account (this is caused by an alkalinization due to
the production of ammonia from peptone).
- Record the results on the result sheet.
bioMérieux SA English - 2
api® 50 CHB/E Medium 07964G - en - 2011/07
bioMérieux SA English - 3
api® 50 CHB/E Medium 07964G - en - 2011/07
QUALITY CONTROL
The media and strips are systematically controlled at various stages of their manufacture. For those users who wish to
perform their own quality control tests with the strip, the following strains may be used :
®
For Bacillus : Paenibacillus polymyxa ATCC 43865
0 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49
24 - + - - + + + - - V + + + + - V - - + - - + - + + + + + + + + + + - + + + + - + + - - - - - - - - -
48 - + - - + + + - - + + + + + - V - - + - - + - + + + + + + + + + + V + + + + - + + - - - - - - - - -
Results obtained after incubation at 30°C.
For Enterobacteriaceae : preferably 1. Klebsiella pneumoniae ssp pneumoniae ATCC 35657 or else :
2. Providencia alcalifaciens ATCC 9886
0 0 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49
1. 24 – + – – + + + – + – + + + + – + – + + + – + + – + + + + + + + + + – – + + – – + – – – – + + – + + –
48 – + – V + + + – + – + + + + – + – + + + – + + V + + + + + + + + + – – + + – – + V – – – + + – + + –
2. 24 – – – – – + – – + – – + + + – – – – – – – – + – – – – – – – – – – – – – – – – – – – – – – – V + – –
48 – V – – – + – – + – – + + + – – – – – – – – + – – – – – – – – – – – – – – – – – – – – – – – + + – –
ATCC : American Type Culture Collection, 10801 University Boulevard, Manassas, VA 20110-2209, USA.
It is the responsibility of the user to perform Quality Control in accordance with any local applicable regulations.
LIMITATIONS OF THE METHOD For Enterobacteriaceae and Vibrionaceae
The API 50 CHB/E system is intended uniquely for the 2930 collection strains and strains of various origins
identification of those species included in the database belonging to species included in the database were
(see Identification Tables at the end of this package tested :
insert). It cannot be used to identify any other - 93.93% of the strains were correctly identified (with or
microorganisms or to exclude their presence. without supplementary tests).
Only pure cultures of a single organism should be used. - 4.47% of the strains were not identified.
- 1.60% of the strains were misidentified.
RANGE OF EXPECTED RESULTS
Consult the Identification Tables at the end of this WASTE DISPOSAL
package insert for the range of expected results for the Dispose of used or unused reagents as well as any other
various biochemical reactions. contaminated disposable materials following procedures
for infectious or potentially infectious products.
PERFORMANCE
It is the responsibility of each laboratory to handle waste
For Bacillus and related genera and effluents produced according to their type and degree
1378 collection strains and strains of various origins of hazardousness and to treat and dispose of them (or
belonging to species included in the database were have them treated and disposed of) in accordance with
tested : any applicable regulations.
- 91.1% of the strains were correctly identified (with or
without supplementary tests). WARRANTY
- 3.9% of the strains were not identified. bioMérieux disclaims all warranties, express or implied,
- 5.0% of the strains were misidentified. including any implied warranties of MERCHANTABILITY
AND FITNESS FOR A PARTICULAR USE. bioMérieux
shall not be liable for any incidental or consequential
damages. IN NO EVENT SHALL BIOMERIEUX’S
LIABLITY TO CUSTOMER UNDER ANY CLAIM
EXCEED A REFUND OF THE AMOUNT PAID TO
BIOMERIEUX FOR THE PRODUCT OR SERVICE
WHICH IS THE SUBJECT OF THE CLAIM.
PROCEDURES p. I
IDENTIFICATION TABLES p. III
LITERATURE REFERENCES p. VI
INDEX OF SYMBOLS p. VII
bioMérieux, the blue logo, API, ATB and apiweb are used, pending and/or registered trademarks belonging to bioMérieux SA or one of its subsidiaries.
CLSI is a trademark belonging to Clinical and Laboratory Standards Institute, Inc.
ATCC is a trademark belonging to American Type Culture Collection.
Any other name or trademark is the property of its respective owner.
Toute la culture
All the culture
Alle Keime
Todo el cultivo
S Tutta la coltura
Toda a cultura
Eau physiologique stérile
Sterile saline Hela kulturen
Sterile Kochsalzlösung Alle bakterier
Agua fisiológica estéril Ca o hodowli
Soluzione fisiologica sterile
Soro fisiológico estéril
2n
2 McF
API NaCl 0.85 % API 50 CHB/E API NaCl 0.85 %
Medium 5 ml Medium Medium 5 ml
facultatif
facultative
fakultativ
facultativo
facoltativo
bioMérieux SA I
api® 50 CHB/E Medium 07964G - xl - 2011/07
Enterobacteriaceae, Vibrionaceae
ou / or / oder / o / / eller /
lub
1
colonie
colony
Kolonie
colonia
colónia
4 McF
koloni
kolonia
API NaCl 0.85 % Eau distillée
Distilled water
Medium
Aqua dest.
5 ml Aqua destilada
ou / or / oder / o / / eller /
lub
Acqua distillata
Água destilada
API Suspension
Medium Destillerat vatten
5 ml Destileret vand
Woda destylowana
1 ml
0.5 McF
API 20 E
API 50 CH
24:00 ± 2:00
36°C ± 2°C
48:00 ± 6:00
TDA : TDA
+ - + - + - IND
VP
: JAMES (IND)
: VP 1 + VP 2
GLU (NO2) : NIT 1 + NIT 2 (+Zn)
API 20 E
bioMérieux SA II
api® 50 CHB/E Medium 07964G - xl - 2011/07
TABLEAU D'IDENTIFICATION / IDENTIFICATION TABLE / PROZENTTABELLE / TABLA DE IDENTIFICACION / TABELLA DI IDENTIFICAZIONE / QUADRO DE IDENTIFICAÇÃO /
/ IDENTIFIERINGSTABELL / IDENTIFIKATIONSTABEL / TABELA IDENTYFIKACYJNA
Bacillus
% de réactions positives dans les conditions d'incubation précisées à droite du tableau / % of positive reactions in the incubation conditions specified on the right-hand side of the table /
% der positiven Reaktionen unter den in der Tabelle angegebenen Inkubationsbedingungen / % de las reacciones positivas en las condiciones de incubación indicadas a continuación /
% di reazioni positive nelle condizioni di incubazione sotto indicate / % das reacções positivas nas condições de incubação indicadas aqui abaixo /
% /% positiva reaktioner under de inkubationsförhållanden som anges i tabellens högra kant /
% positive reaktioner under inkubationsbetingelser som specificeret i tabellens højre side / % pozytywnych reakcji po inkubacji w warunkach okre lonych po prawej stronie tabeli
0 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 API 20 E
API 50 CHB V4.0
0 GLY ERY DARA LARA RIB DXYL LXYL ADO MDX GAL GLU FRU MNE SBE RHA DUL INO MAN SOR MDM MDG NAG AMY ARB ESC SAL CEL MAL LAC MEL SAC TRE INU MLZ RAF AMD GLYG XLT GEN TUR LYX TAG DFUC LFUC DARL LARL GNT 2KG 5KG ONPG ADH LDC ODC CIT H2S URE TDA IND VP GEL NIT TEMP INCUB
Paenibacillus amylolyticus 0 26 0 0 84 84 100 0 0 93 100 100 100 100 0 53 0 1 99 0 46 100 93 100 100 100 100 100 100 100 100 100 100 6 53 100 100 100 0 100 100 0 0 0 6 0 0 0 0 0 99 1 1 1 1 1 8 1 1 30 2 69 29° 48 h
Paenibacillus glucanolyticus 0 61 0 38 100 100 99 11 0 72 88 88 100 88 0 29 11 14 94 11 20 94 100 100 100 100 100 100 100 100 100 100 100 55 75 100 99 99 5 100 100 0 0 0 27 0 0 27 0 1 91 1 1 1 1 1 5 1 1 1 12 35 29° 48 h
Paenibacillus lautus 0 73 0 46 100 93 100 0 26 66 100 100 100 99 0 0 6 6 99 6 6 98 100 100 100 100 100 100 100 100 100 100 98 33 53 100 99 93 6 100 97 0 6 0 40 6 0 53 0 0 99 7 1 1 1 1 7 1 1 64 1 8 29° 48 h
Paenibacillus macerans 0 77 0 51 99 82 99 1 1 82 100 100 100 99 0 58 0 11 93 22 22 77 40 97 99 100 97 99 100 100 100 100 100 88 62 99 99 99 5 97 93 1 0 0 37 51 0 74 1 3 93 1 1 1 1 1 1 1 1 76 30 12 29° 48 h
Paenibacillus polymyxa 0 83 0 2 93 100 97 0 0 83 97 100 99 97 0 2 0 1 99 0 6 71 46 100 100 100 100 99 100 97 100 100 100 69 22 99 99 93 0 97 99 0 0 0 2 0 0 35 0 1 97 2 1 1 2 6 1 1 1 56 70 37 29° 48 h
Paenibacillus validus 0 93 0 0 6 100 93 0 0 6 100 100 100 43 0 0 85 100 100 6 0 100 0 0 0 100 0 25 100 6 25 100 100 50 0 43 68 68 6 6 100 0 25 0 0 6 0 0 0 0 0 0 0 0 0 0 0 0 0 98 0 62 29° 48 h
Virgibacillus pantothenticus 0 55 0 44 0 88 0 0 0 0 98 100 100 98 0 79 0 20 0 50 1 100 100 88 100 100 100 38 100 20 0 94 100 0 0 0 98 5 0 27 66 0 98 0 61 0 0 33 0 0 22 5 1 1 29 12 1 1 1 1 70 25 29° 48 h
* Bacillus non réactif / Bacillus non reactive / Bacillus nicht reaktiv / Bacillus no reactivo / Bacillus non reattivo / Bacillus não reactivo / Bacillus / icke reaktiv Bacillus /
Bacillus ikke reaktiv / Bacillus nie daj cy reakcji = Bacillus sphaericus / fusiformis / badius
** Brevibacillus non réactif / Brevibacillus non reactive / Brevibacillus nicht reaktiv / Brevibacillus no reactivo / Brevibacillus non reattivo / Brevibacillus não reactivo / Brevibacillus /
icke reaktiv Brevibacillus / Brevibacillus ikke reaktiv / Brevibacillus nie daj cy reakcji = Brevibacillus choshinensis / centrosporus / borstelensis / brevis
bioMérieux SA III
api® 50 CHB/E Medium 07964G - xl - 2011/07
TABLEAU D'IDENTIFICATION / IDENTIFICATION TABLE / PROZENTTABELLE / TABLA DE IDENTIFICACION / TABELLA DI IDENTIFICAZIONE / QUADRO DE IDENTIFICAÇÃO /
/ IDENTIFIERINGSTABELL / IDENTIFIKATIONSTABEL / TABELA IDENTYFIKACYJNA
Enterobacteriaceae
% de réactions positives après 48 H (± 6 H) à 36°C ± 2°C / % of positive reactions after 48 hrs. (± 6 hrs.) at 36°C ± 2°C / % der positiven Reaktionen nach 48 Std. (± 6 Std.) bei 36°C ± 2°C /
% de las reacciones positivas después de 48 H (± 6 H) à 36°C ± 2°C / % di reazioni positive dopo 48 ore (± 6 ore) a 36°C ± 2°C / % das reacções positivas após 48 H (± 6 H) a 36°C ± 2°C /
% 48 (± 6 ) 36°C ± 2°C / % positiva reaktioner efter 48 tim. (± 6 tim.) vid 36°C C ± 2°C / % positive reaktioner efter 48 timer (± 6 timer) ved 36°C ± 2°C /
% pozytywnych reakcji po 48 godzinach (± 6 godz.) w 36°C ± 2°C
bioMérieux SA IV
api® 50 CHB/E Medium 07964G - xl - 2011/07
bioMérieux SA V
api® 50 CHB/E Medium 07964G - xl - 2011/07
Bacillus
et apparentés / and related genera / und verwandte Gattungen / y microorganismos próximos /
e generi affini / e semelhantes / / och närstående släkten / og relaterede genera /
i rodzaje pokrewne
1. FINLEY N., FIELDS M.L. 5. MURRAY P.R., BARON E.J., PFALLER M.A.,
Heat Activation and Heat-induced Dormancy of Bacillus TENOVER F.C., YOLKEN R.H.
stearothermophilus spores. Manual of Clinical Microbiology.
(1962) Appl. Microbiol., 10, 231-236. 7th Edition.
2. LE MINOR L., VERON M. (1999) American Society for Microbiology, Washington, D.C.
Bactériologie Médicale. 6. PRETORIUS I.S., DE KOCK M.J., BRITZ T.J.,
2ème édition. POTGIETER H.J. and LATEGAN P.M.
(1989) Flammarion Médecine Sciences. Numerical Taxonomy of alpha-amylase producing Bacillus
3. LOGAN N.A., BERKELEY R.C.W. species.
Identification of Bacillus strains Using the API System. (1986) J. Appl. Bacteriol., 60, 351-360.
(1984) J. Gen. Microbiol., 130, 1871-1882. 7. SELDIN L., PENIDO E.G.
4. LOGAN N.A., CARMAN J.A., MELLING J. and Identification of Bacillus azotofixans using API Tests.
BERKELEY R.C.W. (1986) Antonie van Leeuwenhoek, 52, 403-409.
Identification of Bacillus anthracis by API Tests. 8. SNEATH P.H.A., MAIR N.S., SHARPE E., HOLT J.G.
(1985) J. Med. Microbiol., 20, 75-85. Bergey's Manual of Systematic Bacteriology.
(1986) Williams and Wilkins - Vol. 2.
Enterobacteriaceae + Vibrionaceae
1. BRISOU B., RICHARD C., VIEU J.F., BUISSIERE J. 8. MERGAERT J., VERDONCK L., KERSTERS K., SWINGS J.,
Comparaison de Souches de Salmonella typhimurium isolées BOEUFGRAS J.M., DE LEY J.
chez l'Homme et chez le Pigeon dans la Région Toulonnaise. Numerical Taxonomy of Erwinia Species Using API Systems.
(1975) Med. Mal. Infect. 5, 554-556 (1984) J. Gen. Microbiol, 130, 1893-1910
2. CHOUTEAU J., VIEU J.F., BRAULT G. 9. MURRAY P.R., BARON E.J., PFALLER M.A.,
Epidémiologie de l'Infection Hospitalière à Providencia dans TENOVER F.C., YOLKEN R.H.
un Hôpital Général. Manual of Clinical Microbiology.
(1974) Med. Mal. Infect. 4, 575-578. Seventh Edition.
3. DESCAMPS P., VERON M., LE MINOR S., BUISSIERE J. (1999) American Society for Microbiology, Washington, D.C.
Phénotypes et Marqueurs Epidémiologiques de Salmonella 10. RICHARD C., POPOFF M., PRATS PASTOR G.
typhimurium. Etude Bactériologique d'Infections Urinaires Intra-
(1982) Rev. Epidem. et Santé Publ. 30, 423-435 hospitalières à Proteus rettgeri Fermentant le Lactose.
4. GAVINI F., IZARD D., LECLERC H., DESMONCEAUX M., (1974) Ann. Biol. Clin. 32, 149-154
GAYRAL J.P. 11. VERON M.
Carbon Sources Assimilation Tests: Comparison Between a Nutrition et Taxonomie des Enterobacteriaceae et Bactéries
Conventional Method and a Microtechnic (API), in Study of Voisines.
Enterobacteriaceae. I. Méthode d'Etude des Auxanogrammes.
(1980) Zbl. Bakt. Hyg., I Abt. Orig. C 1, 182-187 (1975) Ann. Microbiol. (Inst. Past.) 126 A, 267-274.
5. GOOR M., MERGAERT J., VERDONCK L., RIJCKAERT C., 12. VERON M., LE MINOR L.
VAN TOMME R., SWINGS J., KERSTERS K., DE LEY J. Nutrition et Taxonomie des Enterobacteriaceae et Bactéries
The Use of API Systems in the Identification of Voisines.
Phytopathogenic Bacteria. II. Résultats d'Ensemble et Classification.
(1984) Med. Fac. Landbouww. Rijksuniv. Gent, 49, 499-507 (1975) Ann. Microbiol. (Inst. Past.) 126 B, 111-123
6. KRIEG N.R., HOLT J.G. 13. VERON M., LE MINOR L.
Bergey’s Manual of Systematic Bacteriology. Nutrition et Taxonomie des Enterobacteriaceae et Bactéries
(1984) Williams and Wilkins – Vol 1. Voisines.
7. LE MINOR L., VERON M. III. Caractères Nutritionnels et Différenciation des Groupes
Bactériologie Médicale. Taxonomiques.
2ème édition. (1975) Ann. Microbiol. (Inst. Past.), 126 B, 125-147.
(1989) Flammarion Médecine Sciences.
bioMérieux SA VI
api® 50 CHB/E Medium 07964G - xl - 2011/07
®
50 CHB/E Medium IVD
bioMérieux SA Português - 1
api® 50 CHB/E Medium 07964G - pt - 2011/07
bioMérieux SA Português - 2
api® 50 CHB/E Medium 07964G - pt - 2011/07
bioMérieux SA Português - 3
api® 50 CHB/E Medium 07964G - pt - 2011/07
CONTROLO DE QUALIDADE
Os meios e galerias são sujeitos a controlos de qualidade sistemáticos nas diferentes etapas do seu fabrico. Além disso,
o utilizador pode efectuar um controlo bacteriológico dos testes da galeria :
Para Bacillus : com a estirpe/cepa Paenibacillus polymyxa ATCC 43865
0 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49
24 - + - - + + + - - V + + + + - V - - + - - + - + + + + + + + + + + - + + + + - + + - - - - - - - - -
48 - + - - + + + - - + + + + + - V - - + - - + - + + + + + + + + + + V + + + + - + + - - - - - - - - -
0 0 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49
1. 24 – + – – + + + – + – + + + + – + – + + + – + + – + + + + + + + + + – – + + – – + – – – – + + – + + –
48 – + – V + + + – + – + + + + – + – + + + – + + V + + + + + + + + + – – + + – – + V – – – + + – + + –
2. 24 – – – – – + – – + – – + + + – – – – – – – – + – – – – – – – – – – – – – – – – – – – – – – – V + – –
48 – V – – – + – – + – – + + + – – – – – – – – + – – – – – – – – – – – – – – – – – – – – – – – + + – –
ATCC : American Type Culture Collection, 10801 University Boulevard, Manassas, VA 20110-2209, USA.
É da responsabilidade do utilizador assegurar que o controlo de qualidade é efectuado em conformidade com a
legislação local em vigor.
LIMITES DO TESTE Para Enterobacteriaceae e Vibrionaceae
O sistema API 50 CHB/E destina-se à identificação das Foram testadas 2930 estirpes/cepas de diversas
espécies presentes na base de dados (consultar os origens e estirpes/cepas de colecção pertencentes às
Quadros de Identificação no final do folheto espécies da base de dados :
informativo), e apenas a estas. Pode ser utilizado para - 93,93% das estirpes/cepas foram correctamente
identificar outros microrganismos ou excluir a sua identificadas (com ou sem testes complementares).
presença. - 4,47% das estirpes/cepas não foram identificadas.
Devem apenas ser utilizadas culturas puras contendo - 1,60% das estirpes/cepas foram mal identificadas.
um único tipo de microrganismo. ELIMINAÇÃO DE RESÍDUOS
RESULTADOS ESPERADOS Eliminar os reagentes utilizados ou não utilizados, bem
Consultar o Quadro de Identificação no final deste folheto como os materiais descartáveis contaminados, em
informativo para saber os resultados esperados para as conformidade com os procedimentos relativos aos
diferentes reacções bioquímicas. produtos infecciosos ou potencialmente infecciosos.
É da responsabilidade de cada laboratório gerir os
COMPORTAMENTO FUNCIONAL resíduos e os efluentes que este produz consoante a sua
Para Bacillus e semelhantes natureza e o seu perigo, e assegurar (ou fazer assegurar)
Foram testadas 1378 estirpes/cepas de diversas o tratamento e a eliminação em conformidade com as
origens e estirpes/cepas de colecção pertencentes às regulamentações aplicáveis.
espécies da base de dados :
- 91,1% das estirpes/cepas foram correctamente
identificadas (com ou sem testes complementares).
- 3,9% das estirpes/cepas não foram identificadas.
- 5,0% das estirpes/cepas foram mal identificadas.
PROCEDIMENTOS p. I
QUADROS DE IDENTIFICAÇÃO p. III
BIBLIOGRAFIA p. VI
QUADRO DE SÍMBOLOS p. VII
A BIOMERIEUX, o logotipo azul, API, ATB e apiweb são marcas utilizadas, depositadas e/ou registadas, propriedade exclusiva da bioMérieux SA ou de uma
das suas filiais.
CLSI é uma marca propriedade exclusiva da Clinical and Laboratory Standards Institute, Inc.
ATCC é uma marca propriedade exclusiva da American Type Culture Collection.
As outras marcas e nomes de produtos mencionados neste documento são marcas comerciais dos seus proprietários respectivos.
Brasil: Distribuído por bioMérieux Brasil, S.A. - Estrada do Mapuá, 491 - Jacarepaguá - R.J. - CEP 22710-261
CNPJ: 33.040.635/0001-71
Atendimento ao Consumidor Tel.: 0800-264848
Prazo de Validade, N° de Lote, N° de Registro de Ministério da Saúde e Responsável Técnico:
VIDE EMBALAGEM
Toute la culture
All the culture
Alle Keime
Todo el cultivo
S Tutta la coltura
Toda a cultura
Eau physiologique stérile
Sterile saline Hela kulturen
Sterile Kochsalzlösung Alle bakterier
Agua fisiológica estéril Ca o hodowli
Soluzione fisiologica sterile
Soro fisiológico estéril
2n
2 McF
API NaCl 0.85 % API 50 CHB/E API NaCl 0.85 %
Medium 5 ml Medium Medium 5 ml
facultatif
facultative
fakultativ
facultativo
facoltativo
bioMérieux SA I
api® 50 CHB/E Medium 07964G - xl - 2011/07
Enterobacteriaceae, Vibrionaceae
ou / or / oder / o / / eller /
lub
1
colonie
colony
Kolonie
colonia
colónia
4 McF
koloni
kolonia
API NaCl 0.85 % Eau distillée
Distilled water
Medium
Aqua dest.
5 ml Aqua destilada
ou / or / oder / o / / eller /
lub
Acqua distillata
Água destilada
API Suspension
Medium Destillerat vatten
5 ml Destileret vand
Woda destylowana
1 ml
0.5 McF
API 20 E
API 50 CH
24:00 ± 2:00
36°C ± 2°C
48:00 ± 6:00
TDA : TDA
+ - + - + - IND
VP
: JAMES (IND)
: VP 1 + VP 2
GLU (NO2) : NIT 1 + NIT 2 (+Zn)
API 20 E
bioMérieux SA II
api® 50 CHB/E Medium 07964G - xl - 2011/07
TABLEAU D'IDENTIFICATION / IDENTIFICATION TABLE / PROZENTTABELLE / TABLA DE IDENTIFICACION / TABELLA DI IDENTIFICAZIONE / QUADRO DE IDENTIFICAÇÃO /
/ IDENTIFIERINGSTABELL / IDENTIFIKATIONSTABEL / TABELA IDENTYFIKACYJNA
Bacillus
% de réactions positives dans les conditions d'incubation précisées à droite du tableau / % of positive reactions in the incubation conditions specified on the right-hand side of the table /
% der positiven Reaktionen unter den in der Tabelle angegebenen Inkubationsbedingungen / % de las reacciones positivas en las condiciones de incubación indicadas a continuación /
% di reazioni positive nelle condizioni di incubazione sotto indicate / % das reacções positivas nas condições de incubação indicadas aqui abaixo /
% /% positiva reaktioner under de inkubationsförhållanden som anges i tabellens högra kant /
% positive reaktioner under inkubationsbetingelser som specificeret i tabellens højre side / % pozytywnych reakcji po inkubacji w warunkach okre lonych po prawej stronie tabeli
0 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 API 20 E
API 50 CHB V4.0
0 GLY ERY DARA LARA RIB DXYL LXYL ADO MDX GAL GLU FRU MNE SBE RHA DUL INO MAN SOR MDM MDG NAG AMY ARB ESC SAL CEL MAL LAC MEL SAC TRE INU MLZ RAF AMD GLYG XLT GEN TUR LYX TAG DFUC LFUC DARL LARL GNT 2KG 5KG ONPG ADH LDC ODC CIT H2S URE TDA IND VP GEL NIT TEMP INCUB
Paenibacillus amylolyticus 0 26 0 0 84 84 100 0 0 93 100 100 100 100 0 53 0 1 99 0 46 100 93 100 100 100 100 100 100 100 100 100 100 6 53 100 100 100 0 100 100 0 0 0 6 0 0 0 0 0 99 1 1 1 1 1 8 1 1 30 2 69 29° 48 h
Paenibacillus glucanolyticus 0 61 0 38 100 100 99 11 0 72 88 88 100 88 0 29 11 14 94 11 20 94 100 100 100 100 100 100 100 100 100 100 100 55 75 100 99 99 5 100 100 0 0 0 27 0 0 27 0 1 91 1 1 1 1 1 5 1 1 1 12 35 29° 48 h
Paenibacillus lautus 0 73 0 46 100 93 100 0 26 66 100 100 100 99 0 0 6 6 99 6 6 98 100 100 100 100 100 100 100 100 100 100 98 33 53 100 99 93 6 100 97 0 6 0 40 6 0 53 0 0 99 7 1 1 1 1 7 1 1 64 1 8 29° 48 h
Paenibacillus macerans 0 77 0 51 99 82 99 1 1 82 100 100 100 99 0 58 0 11 93 22 22 77 40 97 99 100 97 99 100 100 100 100 100 88 62 99 99 99 5 97 93 1 0 0 37 51 0 74 1 3 93 1 1 1 1 1 1 1 1 76 30 12 29° 48 h
Paenibacillus polymyxa 0 83 0 2 93 100 97 0 0 83 97 100 99 97 0 2 0 1 99 0 6 71 46 100 100 100 100 99 100 97 100 100 100 69 22 99 99 93 0 97 99 0 0 0 2 0 0 35 0 1 97 2 1 1 2 6 1 1 1 56 70 37 29° 48 h
Paenibacillus validus 0 93 0 0 6 100 93 0 0 6 100 100 100 43 0 0 85 100 100 6 0 100 0 0 0 100 0 25 100 6 25 100 100 50 0 43 68 68 6 6 100 0 25 0 0 6 0 0 0 0 0 0 0 0 0 0 0 0 0 98 0 62 29° 48 h
Virgibacillus pantothenticus 0 55 0 44 0 88 0 0 0 0 98 100 100 98 0 79 0 20 0 50 1 100 100 88 100 100 100 38 100 20 0 94 100 0 0 0 98 5 0 27 66 0 98 0 61 0 0 33 0 0 22 5 1 1 29 12 1 1 1 1 70 25 29° 48 h
* Bacillus non réactif / Bacillus non reactive / Bacillus nicht reaktiv / Bacillus no reactivo / Bacillus non reattivo / Bacillus não reactivo / Bacillus / icke reaktiv Bacillus /
Bacillus ikke reaktiv / Bacillus nie daj cy reakcji = Bacillus sphaericus / fusiformis / badius
** Brevibacillus non réactif / Brevibacillus non reactive / Brevibacillus nicht reaktiv / Brevibacillus no reactivo / Brevibacillus non reattivo / Brevibacillus não reactivo / Brevibacillus /
icke reaktiv Brevibacillus / Brevibacillus ikke reaktiv / Brevibacillus nie daj cy reakcji = Brevibacillus choshinensis / centrosporus / borstelensis / brevis
bioMérieux SA III
api® 50 CHB/E Medium 07964G - xl - 2011/07
TABLEAU D'IDENTIFICATION / IDENTIFICATION TABLE / PROZENTTABELLE / TABLA DE IDENTIFICACION / TABELLA DI IDENTIFICAZIONE / QUADRO DE IDENTIFICAÇÃO /
/ IDENTIFIERINGSTABELL / IDENTIFIKATIONSTABEL / TABELA IDENTYFIKACYJNA
Enterobacteriaceae
% de réactions positives après 48 H (± 6 H) à 36°C ± 2°C / % of positive reactions after 48 hrs. (± 6 hrs.) at 36°C ± 2°C / % der positiven Reaktionen nach 48 Std. (± 6 Std.) bei 36°C ± 2°C /
% de las reacciones positivas después de 48 H (± 6 H) à 36°C ± 2°C / % di reazioni positive dopo 48 ore (± 6 ore) a 36°C ± 2°C / % das reacções positivas após 48 H (± 6 H) a 36°C ± 2°C /
% 48 (± 6 ) 36°C ± 2°C / % positiva reaktioner efter 48 tim. (± 6 tim.) vid 36°C C ± 2°C / % positive reaktioner efter 48 timer (± 6 timer) ved 36°C ± 2°C /
% pozytywnych reakcji po 48 godzinach (± 6 godz.) w 36°C ± 2°C
bioMérieux SA IV
api® 50 CHB/E Medium 07964G - xl - 2011/07
bioMérieux SA V
api® 50 CHB/E Medium 07964G - xl - 2011/07
Bacillus
et apparentés / and related genera / und verwandte Gattungen / y microorganismos próximos /
e generi affini / e semelhantes / / och närstående släkten / og relaterede genera /
i rodzaje pokrewne
1. FINLEY N., FIELDS M.L. 5. MURRAY P.R., BARON E.J., PFALLER M.A.,
Heat Activation and Heat-induced Dormancy of Bacillus TENOVER F.C., YOLKEN R.H.
stearothermophilus spores. Manual of Clinical Microbiology.
(1962) Appl. Microbiol., 10, 231-236. 7th Edition.
2. LE MINOR L., VERON M. (1999) American Society for Microbiology, Washington, D.C.
Bactériologie Médicale. 6. PRETORIUS I.S., DE KOCK M.J., BRITZ T.J.,
2ème édition. POTGIETER H.J. and LATEGAN P.M.
(1989) Flammarion Médecine Sciences. Numerical Taxonomy of alpha-amylase producing Bacillus
3. LOGAN N.A., BERKELEY R.C.W. species.
Identification of Bacillus strains Using the API System. (1986) J. Appl. Bacteriol., 60, 351-360.
(1984) J. Gen. Microbiol., 130, 1871-1882. 7. SELDIN L., PENIDO E.G.
4. LOGAN N.A., CARMAN J.A., MELLING J. and Identification of Bacillus azotofixans using API Tests.
BERKELEY R.C.W. (1986) Antonie van Leeuwenhoek, 52, 403-409.
Identification of Bacillus anthracis by API Tests. 8. SNEATH P.H.A., MAIR N.S., SHARPE E., HOLT J.G.
(1985) J. Med. Microbiol., 20, 75-85. Bergey's Manual of Systematic Bacteriology.
(1986) Williams and Wilkins - Vol. 2.
Enterobacteriaceae + Vibrionaceae
1. BRISOU B., RICHARD C., VIEU J.F., BUISSIERE J. 8. MERGAERT J., VERDONCK L., KERSTERS K., SWINGS J.,
Comparaison de Souches de Salmonella typhimurium isolées BOEUFGRAS J.M., DE LEY J.
chez l'Homme et chez le Pigeon dans la Région Toulonnaise. Numerical Taxonomy of Erwinia Species Using API Systems.
(1975) Med. Mal. Infect. 5, 554-556 (1984) J. Gen. Microbiol, 130, 1893-1910
2. CHOUTEAU J., VIEU J.F., BRAULT G. 9. MURRAY P.R., BARON E.J., PFALLER M.A.,
Epidémiologie de l'Infection Hospitalière à Providencia dans TENOVER F.C., YOLKEN R.H.
un Hôpital Général. Manual of Clinical Microbiology.
(1974) Med. Mal. Infect. 4, 575-578. Seventh Edition.
3. DESCAMPS P., VERON M., LE MINOR S., BUISSIERE J. (1999) American Society for Microbiology, Washington, D.C.
Phénotypes et Marqueurs Epidémiologiques de Salmonella 10. RICHARD C., POPOFF M., PRATS PASTOR G.
typhimurium. Etude Bactériologique d'Infections Urinaires Intra-
(1982) Rev. Epidem. et Santé Publ. 30, 423-435 hospitalières à Proteus rettgeri Fermentant le Lactose.
4. GAVINI F., IZARD D., LECLERC H., DESMONCEAUX M., (1974) Ann. Biol. Clin. 32, 149-154
GAYRAL J.P. 11. VERON M.
Carbon Sources Assimilation Tests: Comparison Between a Nutrition et Taxonomie des Enterobacteriaceae et Bactéries
Conventional Method and a Microtechnic (API), in Study of Voisines.
Enterobacteriaceae. I. Méthode d'Etude des Auxanogrammes.
(1980) Zbl. Bakt. Hyg., I Abt. Orig. C 1, 182-187 (1975) Ann. Microbiol. (Inst. Past.) 126 A, 267-274.
5. GOOR M., MERGAERT J., VERDONCK L., RIJCKAERT C., 12. VERON M., LE MINOR L.
VAN TOMME R., SWINGS J., KERSTERS K., DE LEY J. Nutrition et Taxonomie des Enterobacteriaceae et Bactéries
The Use of API Systems in the Identification of Voisines.
Phytopathogenic Bacteria. II. Résultats d'Ensemble et Classification.
(1984) Med. Fac. Landbouww. Rijksuniv. Gent, 49, 499-507 (1975) Ann. Microbiol. (Inst. Past.) 126 B, 111-123
6. KRIEG N.R., HOLT J.G. 13. VERON M., LE MINOR L.
Bergey’s Manual of Systematic Bacteriology. Nutrition et Taxonomie des Enterobacteriaceae et Bactéries
(1984) Williams and Wilkins – Vol 1. Voisines.
7. LE MINOR L., VERON M. III. Caractères Nutritionnels et Différenciation des Groupes
Bactériologie Médicale. Taxonomiques.
2ème édition. (1975) Ann. Microbiol. (Inst. Past.), 126 B, 125-147.
(1989) Flammarion Médecine Sciences.
bioMérieux SA VI
api® 50 CHB/E Medium 07964G - xl - 2011/07
PW3
ISOLATION AND IDENTIFICATION OF ENTEROBACTERIACEAE STRAINS
Master in Microbiology
1. Isolation
1.1 For each supplied sample prepare, under aseptic conditions, a 10% suspension (% w/v
or % v/v depending on the type of sample) in 10 ml of saline solution (0.9% NaCl) or
peptonated water (0.1% peptone).
If necessary, fragment, grind or macerate the sample in advance for homogenisation.
1.2 After vortexing, inoculate an aliquot of each sample onto a Mac Conkey medium plate.
To obtain isolated colonies, dilute previously (10-2) and plate 0.1 ml or alternatively
inoculate a loop of the original suspension.
1.4 Select two colonies which, given the characteristics described for this medium (see
Annex), may belong to different taxa.
Confirm that the bacilli are gram-negative, oxidase-negative.
1.5 Isolate each of the selected strains in pure culture by successive passages (at least 3)
by non-selective means (nutritive agar, NA). Between each passage, incubate the plates at
37ºC.
1.6. At the end of the isolation, confirm that the bacilli are gram negative and oxidase
negative. Refer to the strains with code ENTxA and ENTxB where x is the student group
number.
2. Identification
2.1. For the strain of your group that is selected in the class, carry out its identification at
species level, using the miniaturized galleries API 20E, according to the manufacturer's
instructions.
2.2 If it is impossible to achieve an identification, check that the final isolation plate does
not have a mixed culture (failure of the purification process).
3.1. inoculate the individual strains in the supplied differential media (at least 2 strains per
plate).
The list of media to be used may include: EMB, Mac Conkey, Endo, Chromocult Coliform
agar and Rambach agar.
3.3 Observe the plates after incubation and compare the results obtained in the
identification with the observed differential characteristics.
Meios selectivos e diferenciais para Enterobacteriaceae (1)
triptona
fermentação da lactose
BCPLA
extracto de carne indicador de pH (púrpura de meio não selectivo para
Bromocresol - detecção e isolamento de
Purple Lactose
lactose bromocresola) permite
púrpura de bromocresol detectar enterobacteriácias
Agar agar
a produção de ácido
hidrol. pancreático de gelatina
hidrolisado parcial de carne
triptona violeta cristal inibe
fermentação da lactose meio selectivo para
lactose crescimento bactérias
indicador de pH (vermelho isolamento de Salmonella,
MacConkey sais biliares Gram + e sais biliares
neutro) permite detectar a Shigella e bactérias
cloreto de sódio inibem crescimento outras
vermelho neutro produção de ácido coliformes
bactérias Gram -
violeta cristal
agar
peptona
EMB fosfato dipotássico fermentação da lactose meio selectivo para
lactose eosina-y e azul de metileno
Eosin Methylene- revelada pela precipitação detecção e isolamento de
sucrose inibem crescimento de
Blue Lactose dos corantes sob as colónias enterobacteriácias
eosina-y (corante) bactérias Gram +
Sucrose Agar azul de metileno
patogénicas
agar
hidrol. pancreático de carne fermentação da lactose
fosfato dipotássico
sulfito de sódio e fucsina revelada pela reacção de um meio selectivo para
lactose
Endo sulfito de sódio
inibem crescimento de intermediário metabólico detecção e isolamento de
fucsina básica (corante) bactérias Gram + (acetaldeído) com a fucsina bactérias coliformes
agar sulfatada
peptona
cloreto de sódio o carácter diferencial
di-hidrogenofosfato de sódio baseia-se na utilização
produção de
hidrogenofosfato dissódico (ou não) de 2 substratos
tergitol ß-galactosidases
piruvato de sódio cromogéneosb :
Chromocult inibe Gram + e/ou ß-glucuronidases
triptofano X-glucoronato(ß-
e algumas Gram - produção de indol
sorbitol glucuronidase)
tergitol (detergente) (degradação do triptofanoa) e Salmon-Gal
mistura cromogénea (ß-galactosidase)
agar
o carácter diferencial
baseia-se na utilização
peptona (ou não) de um substratoc
cloreto de sódio produção de cromogéneo para a
desoxicolato de sódio (deterg.) desoxicolato de sódio ß-galactosidases ß-galactosidase e na
Rambach propilenoglicol inibe Gram + fermentação do capacidade de fermentação
mistura cromogénea propilenoglicol do propilenoglicol
agar (a produção de ácido é
detectada por um
indicador de pHd)
a. A degradação (ou não) do triptofano é um teste adicional que pode ser realizado para E. coli (indol +), colocando uma gota do reagente de Kovacs sobre as
colónias. [Resultado positivo: o reagente adquire uma côr vermelha após alguns segundos]
b. A degradação de X-glucoronato produz uma coloração azul claro (a turquesa) enquanto que a degradação de Salmon-Gal, produz uma coloração salmão (a
vermelho). A utilização de ambos os substratos é produz uma coloração azul escuro (a violeta).
c. A degradação do substrato cromogéneo produz uma coloração azul - esverdeada (a azul - violeta).
d. O indicador de pH utilizado para detectar a fermentação do propilenoglicol é vermelho em meio ácido (resultado positivo).
TP3
Enterobacteriaceae
Bactérias Outras
coliformes Enterobacteriaceae
Escherichia coli
Enterobacter Salmonella
Klebsiella Shigella
VRBGA
GLU +
Violet Red Bile
colónias rosa-violeta
Glucose Agar
BCPLA
Bromocresol LAC + LAC -
Purple Lactose colónias amarelas mucosas colónias incolores
Agar
LAC +
colónias LAC + LAC -
MacConkey rosa-vermelhas colónias colónias
com halo sais rosa-vermelhas incolores
biliares precipitados
LAC +
EMB
colónias LAC + LAC -
Eosin Methylene-
vermelhas com colónias colónias
Blue Lactose
brilho metálico vermelhas incolores
Sucrose Agar
verde
LAC +
colónias LAC + LAC -
Endo vermelhas com colónias colónias
brilho metálico vermelhas incolores
verde
ß-galactosidases -
ß-glucuronidases +
colónias
ß-galactosidases + ß-galactosidases + azul claro a turquesa
ß-glucuronidases + ß-glucuronidases -
Chromocult (algumas Salmonella)
Bacteriologia
colónias colónias
azul escuro a violeta salmão a vermelho
ß-galactosidases -
ß-glucuronidases -
colónias incolores
ß-galactosidases +
ß-galactosidases - ß-galactosidases -
não fermenta
fermenta não fermenta
propilenoglicol
Rambach propilenoglicol propilenoglicol
colónias
colónias colónias
azul-esverdeado
vermelhas incolores
a violeta
50 110 07705G - en - 2012/08
®
OF Medium IVD
bioMérieux SA English - 1
API® OF Medium 07705G - en - 2012/08
BIOMERIEUX, the blue logo, API and API logo are used, pending and/or registered trademarks belonging to bioMérieux, or one of its subsidiaries, or one of its
companies.
CLSI is a trademark belonging to Clinical Laboratory and Standards Institute, Inc.
The ATCC trademark and trade name and any and all ATCC catalog numbers are trademarks of the American Type Culture Collection.
Any other name or trademark is the property of its respective owner.
bioMérieux SA I
API® OF Medium 07705G - xl - 2012/08
Se handhavandebeskrivningen / Se brugsanvisning
Sprawd w instrukcji obs ugi
Contenu suffisant pour "n" tests
Contains sufficient for <n> tests
Inhalt ausreichend für <n> Prüfungen
Contenido suficiente para <n> ensayos
Contenuto sufficiente per "n" saggi
Conteúdo suficiente para “n” ensaios
« »
Räcker till "n" antal tester
Indeholder tilstrækkeligt til "n" test
Wystarczy na wykonanie <n> testów
®
OF Medium IVD
bioMérieux SA Português - 1
API® OF Medium 07705G - pt - 2012/08
Brasil: Distribuído por bioMérieux Brasil, S.A. - Estrada do Mapuá, 491 - Jacarepaguá - R.J. - CEP 22710-261
CNPJ: 33.040.635/0001-71
Atendimento ao Consumidor Tel.: 0800-264848
Prazo de Validade, N° de Lote, N° de Registro de Ministério da Saúde e Responsável Técnico:
VIDE EMBALAGEM
bioMérieux SA I
API® OF Medium 07705G - xl - 2012/08
Se handhavandebeskrivningen / Se brugsanvisning
Sprawd w instrukcji obs ugi
Contenu suffisant pour "n" tests
Contains sufficient for <n> tests
Inhalt ausreichend für <n> Prüfungen
Contenido suficiente para <n> ensayos
Contenuto sufficiente per "n" saggi
Conteúdo suficiente para “n” ensaios
« »
Räcker till "n" antal tester
Indeholder tilstrækkeligt til "n" test
Wystarczy na wykonanie <n> testów
®
20 E TM IVD
bioMérieux SA English - 1
api® 20 E TM 07584J - en - 2010/05
bioMérieux SA English - 2
api® 20 E TM 07584J - en - 2010/05
In some cases, the 7-digit profile is not discriminatory - Motility (MOB) : Inoculate an ampule of API M Medium
enough and the following supplementary tests need to be (see package insert).
carried out : - Growth on MacConkey agar medium (McC) : Streak a
- Reduction of nitrates to nitrites (NO2) and N2 gas (N2) : MacConkey agar plate (see package insert).
add 1 drop each of NIT 1 and NIT 2 reagents to the GLU - Oxidation of glucose (OF-O) : Inoculate an ampule of
tube. Wait 2 to 5 minutes. A red color indicates a API OF Medium (see package insert).
positive reaction (NO2). A negative reaction (yellow) - Fermentation of glucose (OF-F) : Inoculate an ampule of
may be due to the reduction to nitrogen (as sometimes API OF Medium (see package insert).
evidenced by gas bubbles) : add 2 to 3 mg of Zn These supplementary tests, indicated in the introduction
reagent to the GLU tube. After 5 minutes, if the tube section (Profile coding) of the Analytical Profile Index, may
remains yellow this indicates a positive reaction (N2) to be used to form a 9-digit profile. Identification is then
be recorded on the result sheet. If the test turns orange- obtained using the identification software.
red, this is a negative reaction : the nitrates still present
in the tube have been reduced by the Zinc.
This reaction is useful when testing Gram-negative,
oxidase positive rods.
NOTE : For the same reason as the indole test (see the 5 315 173 (57) Enterobacter gergoviae
note in the paragraph "Reading the strip"), the nitrate Further tests may be proposed in case of low
reduction test must be performed last. discrimination. Refer to the identification software or
Analytical Profile Index.
QUALITY CONTROL
The media, strips and reagents are systematically quality controlled at various stages of their manufacture.
Streamlined quality control may be used to confirm acceptable performance of the API 20 E system after
shipping/storage. This methodology may be performed by following the instructions above for testing and meeting the
®
criteria stated in CLSI M50-A Quality Control for Commercial Microbial Identification Systems.
Testing may be conducted using Proteus mirabilis ATCC® 35659 to evaluate the performance of the ODC and ARA
tests. Tests performed by bioMérieux has shown that the ODC and ARA tests are the most labile on the API 20 E strip.
When testing the strip, Proteus mirabilis ATCC 35659 can be used to detect degradation.
For those users who are required to perform comprehensive quality control testing with the strip, the following five
strains should be tested to demonstrate positive and negative reactivity for most of the API 20 E tests.
1. Proteus mirabilis ATCC 35659 4. Escherichia coli ATCC 25922
2. Stenotrophomonas maltophilia ATCC 51331 5. Klebsiella pneumoniae ssp pneumoniae ATCC 35657
3. Enterobacter cloacae ATCC 13047
ATCC : American Type Culture Collection, 10801 University Boulevard, Manassas, VA 20110-2209, USA.
ONPG ADH LDC ODC CIT H2S URE TDA IND VP GEL GLU MAN INO SOR RHA SAC MEL AMY ARA NO2 N2*
1. – – – + V + + + – – V + – – – – V – – – + –
2. + – V – V – – – – – + – – – – – – – – – – –
3. + + – V + – – – – + – + + V + + + + + + + –
4. + – + + – – – – + – – + + – + + – + – + + –
5. + – + – + – V – – V – + + + + + + + + + + –
* The N2 (+) state may be observed for the strains ATCC 13047, ATCC 25922 and ATCC 35657.
Profile obtained after 24-48 hours of incubation for the strain ATCC 51331, using colonies grown on Trypticase Soy agar + blood.
Profiles obtained after 18-24 hours of incubation for the other strains, using colonies grown on Trypticase Soy agar + blood.
Bacterial suspensions prepared in API NaCl 0.85 % Medium.
It is the responsibility of the user to perform Quality Control in accordance with any local applicable regulations.
bioMérieux SA English - 3
api® 20 E TM 07584J - en - 2010/05
PROCEDURE p. I
IDENTIFICATION TABLE p. II
READING TABLE p. IV
SUPPLEMENTARY TESTS p. VII
LITERATURE REFERENCES p. VIII
INDEX OF SYMBOLS p. IX
BIOMERIEUX, the blue logo, API and apiweb are used, pending and/or registered trademarks belonging to bioMérieux SA or one of its subsidiaries.
CLSI is a trademark belonging to Clinical Laboratory and Standards Institute, Inc.
ATCC is a trademark belonging to American Type Culture Collection.
Any other name or trademark is the property of its respective owner.
OX
- | CIT |
- | VP |
- | GEL |
API 20 E
ADH ODC
H2S - URE
36°C
18:00-24:00 / 48:00 ±
2°C
Tests < 3
(y compris / including /
einschließlich / incluído /
compreso / incluindo / API 20 E
/ inklusive /
inklusiv / w czaj c test GLU) TDA : TDA
IND : JAMES
VP : VP 1 + VP 2
GLU (NO2) : NIT 1 + NIT 2 (+Zn)
API 20 E
+ - + - + -
bioMérieux SA I
api® 20 E TM 07584J - xl - 2010/05
TABLEAU D'IDENTIFICATION / IDENTIFICATION TABLE / PROZENTTABELLE / TABLA DE IDENTIFICACION / TABELLA DI IDENTIFICAZIONE / QUADRO DE IDENTIFICAÇÃO
/ IDENTIFIKATIONSTABEL / IDENTIFIERINGSTABELL / TABELA IDENTYFIKACYJNA
% de réactions positives après 18-24 / 48 h à 36°C ± 2°C / % of positive reactions after 18-24 / 48 hrs. at 36°C ± 2°C / % der positiven Reaktionen nach 18-24 / 48 h bei 36°C ± 2°C /
% de las reacciones positivas después de 18-24 / 48 H a 36°C ± 2°C / % di reazioni positive dopo 18-24 / 48 ore a 36°C ± 2°C / % de reacções positivas após 18-24 / 48 h a 36º C ± 2º C /
% 18-24 / 48 36°C ± 2°C / % positiva reaktioner efter 18-24 / 48 timmar vid 36°C ± 2°C / % af positive reaktioner efter 18-24 / 48 timer ved 36°C ± 2°C /
% pozytywnych reakcji po 18-24 / 48 godzinach w 36°C ± 2°C
API 20 E V4.1 ONPG ADH LDC ODC CIT H2S URE TDA IND VP GEL GLU MAN INO SOR RHA SAC MEL AMY ARA OX NO2 N2 MOB McC OF/O OF/F
Buttiauxella agrestis 100 0 0 85 25 0 0 0 0 0 0 100 100 0 1 99 0 92 99 100 0 100 0 100 100 100 100
Cedecea davisae 99 89 0 99 75 0 0 0 0 89 0 100 100 10 0 0 100 0 100 1 0 99 0 87 100 100 100
Cedecea lapagei 99 99 0 0 75 0 0 0 0 90 0 100 99 0 0 0 0 1 100 1 0 99 0 87 100 100 100
Citrobacter braakii 50 45 0 99 75 81 1 0 4 0 0 100 100 1 100 100 1 91 99 99 0 100 0 95 100 100 100
Citrobacter freundii 90 24 0 0 75 75 1 0 1 0 0 100 99 25 99 99 99 82 40 99 0 98 0 95 100 100 100
Citrobacter koseri/amalonaticus 99 75 0 100 97 0 1 0 99 0 0 100 100 25 99 99 1 1 98 99 0 100 0 95 100 100 100
Citrobacter koseri/farmeri 99 2 0 100 25 0 1 0 99 0 0 100 100 1 99 99 99 80 99 99 0 100 0 95 100 100 100
Citrobacter youngae 100 50 0 1 80 80 0 0 1 0 0 100 100 0 95 100 1 0 25 100 0 85 0 95 100 100 100
Edwardsiella hoshinae 0 0 100 99 50 94 0 0 99 0 0 100 100 0 0 1 100 0 0 1 0 100 0 100 100 100 100
Edwardsiella tarda 0 0 100 99 1 75 0 0 99 0 0 100 0 0 0 0 0 0 0 0 0 100 0 98 100 100 100
Enterobacter aerogenes 99 0 99 98 82 0 1 0 0 85 0 99 99 99 99 99 99 99 99 99 0 100 0 97 100 100 100
Enterobacter amnigenus 1 99 25 0 99 40 0 0 0 0 75 0 100 100 0 1 100 99 99 99 99 0 100 0 92 100 100 100
Enterobacter amnigenus 2 99 80 0 99 80 0 0 0 0 75 0 100 100 0 99 100 1 99 99 99 0 100 0 100 100 100 100
Enterobacter asburiae 100 25 0 99 80 0 0 0 0 10 0 100 99 25 100 0 99 0 100 100 0 100 0 95 100 100 100
Enterobacter cancerogenus 100 75 0 99 99 0 0 0 0 89 0 100 100 0 1 100 1 1 100 100 0 100 0 99 100 100 100
Enterobacter cloacae 98 82 1 92 90 0 1 0 0 85 0 99 99 12 90 85 96 90 99 99 0 100 0 95 100 100 100
Enterobacter gergoviae 99 0 32 100 75 0 99 0 0 90 0 100 99 23 1 100 99 100 99 100 0 100 0 90 100 100 100
Enterobacter intermedius 99 0 0 99 1 0 0 0 0 2 0 100 97 0 88 99 40 100 99 99 0 100 0 92 100 100 100
Enterobacter sakazakii 100 96 0 91 94 0 1 0 25 91 10 100 100 75 8 99 99 99 99 99 0 100 0 96 100 100 100
Escherichia coli 1 90 1 74 70 0 1 3 0 89 0 0 99 98 1 91 82 36 75 3 99 0 100 0 95 100 100 100
Escherichia coli 2 26 1 45 20 0 1 1 0 50 0 0 99 90 1 42 30 3 3 1 70 0 98 0 5 100 100 100
Escherichia fergusonii 96 1 99 100 1 0 0 0 99 0 0 100 99 1 0 87 0 1 99 99 0 100 0 93 100 100 100
Escherichia hermannii 100 0 1 100 1 0 0 0 99 0 0 100 100 0 0 99 25 0 99 99 0 100 0 99 100 100 100
Escherichia vulneris 100 30 50 0 0 0 0 0 0 0 0 100 100 0 1 95 7 95 95 99 0 100 0 100 100 100 100
Ewingella americana 98 0 0 0 75 0 0 0 0 95 1 99 99 0 0 1 0 1 50 1 0 100 0 60 100 100 100
Hafnia alvei 1 75 0 99 98 50 0 10 0 0 50 0 99 99 0 1 99 0 0 25 99 0 100 0 85 100 100 100
Hafnia alvei 2 50 0 99 99 1 0 1 0 0 10 0 99 98 0 1 1 1 0 0 1 0 100 0 0 100 100 100
Klebsiella oxytoca 99 0 80 0 89 0 78 0 99 80 0 100 100 99 100 99 99 100 100 100 0 100 0 0 100 100 100
Klebsiella pneumoniae ssp ozaenae 94 18 25 1 18 0 1 0 0 1 0 99 96 57 66 58 20 80 97 85 0 92 0 0 100 100 100
Klebsiella pneumoniae ssp pneumoniae 99 0 73 0 86 0 75 0 0 90 0 100 99 99 99 99 99 99 99 99 0 100 0 0 100 100 100
Klebsiella pneumoniae ssp rhinoscleromatis 1 0 0 0 0 0 0 0 0 0 0 99 100 90 90 75 75 1 99 10 0 100 0 0 100 100 100
Kluyvera spp 95 0 25 99 60 0 0 0 80 0 0 100 99 0 25 93 89 99 99 99 0 95 0 94 100 100 100
Leclercia adecarboxylata 99 0 0 0 0 0 1 0 99 0 1 100 99 0 2 100 66 99 99 100 0 100 0 100 100 100 100
Moellerella wisconsensis 97 0 0 0 40 0 0 0 15 1 0 100 1 0 0 0 100 99 0 0 0 90 0 0 100 100 100
Morganella morganii 1 0 10 98 1 1 99 93 99 0 0 99 0 0 0 0 1 0 0 0 0 88 0 95 100 100 100
Pantoea spp 1 85 1 0 0 13 0 1 0 1 9 1 100 99 1 26 1 98 26 59 61 0 85 0 85 100 100 100
Pantoea spp 2 99 1 0 0 99 0 1 0 53 62 4 100 99 36 82 90 98 81 99 99 0 85 0 85 100 100 100
Pantoea spp 3 99 1 0 0 21 0 1 0 1 86 15 100 99 34 1 97 93 23 65 97 0 85 0 85 100 100 100
Pantoea spp 4 86 1 0 0 29 0 1 0 59 1 1 99 100 10 32 99 72 89 99 99 0 85 0 85 100 100 100
Proteus mirabilis 1 0 0 99 50 75 99 98 1 1 82 98 0 0 0 0 1 0 0 0 0 93 0 95 100 100 100
Proteus penneri 1 0 0 0 1 20 100 99 0 0 50 99 0 0 0 0 100 0 1 0 0 99 0 85 100 100 100
Proteus vulgaris group 1 0 0 0 12 83 99 99 92 0 74 99 1 1 0 1 89 0 66 1 0 100 0 94 100 100 100
Providencia alcalifaciens/rustigianii 0 0 0 0 80 0 0 100 99 0 0 99 1 1 0 0 1 0 0 1 0 100 0 96 100 100 100
Providencia rettgeri 1 1 0 0 74 0 99 99 90 0 0 98 82 78 1 50 25 0 40 1 0 98 0 94 100 100 100
Providencia stuartii 1 0 0 0 85 0 30 98 95 0 0 98 3 80 0 0 15 0 0 0 0 100 0 85 100 100 100
Rahnella aquatilis 100 0 0 0 50 0 0 1 0 99 0 100 100 0 98 99 100 97 100 98 0 100 0 6 100 100 100
Raoultella ornithinolytica 100 0 99 99 99 0 85 0 100 65 0 100 100 99 100 100 100 100 100 100 0 100 0 0 100 100 100
Raoultella terrigena 100 0 99 6 52 0 0 0 0 75 0 99 99 99 99 99 100 100 100 99 0 100 0 0 100 100 100
Salmonella choleraesuis ssp arizonae 98 75 97 98 75 99 0 0 1 0 0 100 99 0 99 99 1 78 0 99 0 100 0 99 100 100 100
Salmonella choleraesuis ssp choleraesuis 0 15 99 99 6 64 0 0 0 0 0 100 99 0 98 99 0 20 0 0 0 100 0 95 100 100 100
Salmonella ser.Gallinarum 0 1 100 1 0 25 0 0 0 0 0 100 100 0 0 1 0 0 0 100 0 100 0 0 100 100 100
bioMérieux SA II
api® 20 E TM 07584J - xl - 2010/05
API 20 E V4.1 ONPG ADH LDC ODC CIT H2S URE TDA IND VP GEL GLU MAN INO SOR RHA SAC MEL AMY ARA OX NO2 N2 MOB McC OF/O OF/F
Salmonella ser.Paratyphi A 0 5 0 99 0 1 0 0 0 0 0 100 99 0 99 98 0 96 0 99 0 100 0 95 100 100 100
Salmonella ser.Pullorum 0 1 75 100 0 85 0 0 0 0 0 100 100 0 0 100 0 0 0 75 0 100 0 0 100 100 100
Salmonella typhi 0 1 99 0 0 8 0 0 0 0 0 100 99 0 99 0 0 99 0 0 0 100 0 97 100 100 100
Salmonella spp 1 56 82 93 65 83 0 0 1 0 1 99 100 40 99 86 1 90 1 99 1 100 0 95 100 100 100
Serratia ficaria 99 0 0 0 100 0 0 0 0 40 90 100 100 50 99 74 99 99 100 99 0 92 0 100 100 100 100
Serratia fonticola 99 0 73 99 75 0 0 0 0 0 0 100 100 97 100 99 30 99 99 99 0 99 0 91 100 100 100
Serratia liquefaciens 95 1 78 98 80 0 2 0 0 59 65 100 99 80 98 2 99 72 97 97 0 100 0 95 100 100 100
Serratia marcescens 94 0 95 95 96 0 25 0 1 70 87 100 99 85 98 1 99 68 97 25 0 95 0 97 100 100 100
Serratia odorifera 1 95 0 95 99 95 0 0 0 99 50 99 100 99 99 99 99 99 99 99 99 0 99 0 100 100 100 100
Serratia odorifera 2 95 0 96 1 95 0 0 0 99 50 99 100 99 99 99 99 1 99 99 95 0 99 0 100 100 100 100
Serratia plymuthica 99 0 0 0 65 0 0 0 0 65 50 100 90 70 70 1 99 85 98 98 0 99 0 50 100 100 100
Serratia rubidaea 99 0 30 0 92 0 1 0 0 71 82 99 99 75 1 3 99 95 99 99 0 100 0 85 100 100 100
Shigella spp 1 0 0 1 0 0 0 0 29 0 0 99 63 0 7 7 1 20 0 50 0 100 0 0 100 100 100
Shigella sonnei 96 0 0 93 0 0 0 0 0 0 0 99 99 0 1 75 1 1 0 99 0 100 0 0 100 100 100
Yersinia enterocolitica 80 0 0 90 0 0 98 0 50 5 0 99 99 25 98 1 99 4 75 75 0 98 0 2 100 100 100
Yersinia frederiksenii/intermedia 99 0 0 75 1 0 99 0 99 1 0 100 99 25 99 99 99 1 99 99 0 98 0 5 100 100 100
Yersinia kristensenii 80 0 0 80 0 0 99 0 97 0 0 100 99 10 99 0 0 0 99 99 0 98 0 5 100 100 100
Yersinia pestis 68 0 0 0 0 0 0 0 0 1 0 99 99 0 70 0 0 0 30 30 0 47 0 0 99 100 100
Yersinia pseudotuberculosis 98 0 0 0 1 0 99 0 0 0 0 99 97 0 0 75 0 50 25 50 0 95 0 0 100 100 100
Aeromonas hydrophila gr. 1 98 90 25 1 25 0 0 0 85 25 90 99 99 1 3 5 97 1 75 75 100 97 0 95 99 99 99
Aeromonas hydrophila gr. 2 99 97 80 1 80 0 0 0 85 80 97 97 99 9 9 1 80 1 75 5 100 97 0 95 99 99 99
Aeromonas salmonicida ssp salmonicida 1 60 1 0 0 0 0 0 1 0 75 50 54 0 0 0 0 0 1 0 100 98 0 1 99 99 99
Grimontia hollisae 1 0 0 0 0 0 0 0 94 0 0 10 0 0 0 0 0 0 0 0 100 100 0 0 99 99 99
Photobacterium damselae 1 99 75 0 1 0 98 0 0 10 1 50 0 0 0 0 1 0 0 0 100 100 0 25 99 99 99
Plesiomonas shigelloides 95 99 100 100 0 0 0 0 100 0 0 99 0 99 0 0 0 0 0 0 100 99 0 95 99 99 99
Vibrio alginolyticus 0 0 98 75 60 0 1 0 100 10 75 99 100 0 1 0 100 0 10 1 100 47 0 100 99 94 94
Vibrio cholerae 98 1 94 97 75 0 0 0 99 58 92 98 98 0 0 0 94 0 5 0 100 96 0 100 96 99 99
Vibrio fluvialis 95 99 0 0 1 0 0 0 80 0 75 75 80 0 1 0 75 0 36 75 100 100 0 100 99 99 99
Vibrio mimicus 99 0 99 99 50 0 0 0 99 1 99 99 99 0 0 0 0 0 0 0 100 95 0 100 95 99 99
Vibrio parahaemolyticus 0 0 100 99 50 0 1 0 100 1 75 100 99 0 0 1 1 0 12 50 100 63 0 100 98 99 99
Vibrio vulnificus 99 0 91 90 25 0 0 0 99 1 99 99 75 0 0 0 1 0 90 0 99 54 0 100 99 99 99
Pasteurella aerogenes 99 0 0 80 0 0 99 0 0 0 0 99 0 97 0 1 99 0 0 75 75 100 0 0 100 100 100
Pasteurella multocida 1 4 0 0 25 0 0 0 0 99 0 0 29 1 0 1 0 75 0 0 0 99 90 0 0 2 23 23
Pasteurella multocida 2 7 0 0 45 0 0 0 0 99 0 0 44 99 0 99 0 99 0 0 0 89 90 0 0 2 23 23
Pasteurella pneumotropica/ Mannheimia haemolytica 60 0 1 10 0 0 25 0 15 7 3 35 12 12 12 1 35 1 2 1 80 99 0 0 9 33 33
Acinetobacter baumannii/calcoaceticus 0 0 0 0 51 0 1 0 0 5 5 99 0 0 0 0 0 99 1 99 0 3 0 0 90 98 0
Bordetella/Alcaligenes/Moraxella spp * 0 0 0 0 52 0 14 1 0 25 1 0 0 0 0 0 0 0 0 0 95 62 1 75 75 0 0
Burkholderia cepacia 50 0 25 16 78 0 0 0 0 1 43 60 1 0 0 0 13 0 7 20 90 40 0 99 88 97 0
Chromobacterium violaceum 0 99 0 0 75 0 0 0 14 0 99 99 0 0 0 0 10 0 0 0 99 75 0 99 99 99 99
Chryseobacterium indologenes 5 0 0 0 12 0 90 0 75 0 80 0 0 0 0 0 0 0 0 0 99 20 0 0 57 90 10
Chryseobacterium meningosepticum 77 0 0 0 20 0 1 0 85 0 90 0 0 0 0 0 0 0 0 0 99 6 0 0 48 93 6
Eikenella corrodens 0 0 75 99 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 100 95 0 1 1 49 49
Myroides /Chryseobacterium indologenes 0 0 0 0 50 0 75 0 0 1 75 0 0 0 0 0 0 0 0 0 99 0 0 0 84 2 2
Ochrobactrum anthropi 15 0 0 0 30 0 25 1 0 15 0 1 0 0 0 0 0 0 0 10 90 42 60 99 99 47 0
Pseudomonas aeruginosa 0 89 0 0 92 0 25 0 0 1 75 50 0 0 0 0 1 10 1 25 97 12 56 97 100 98 0
Pseudomonas fluorescens/putida 0 75 0 0 75 0 0 0 0 10 27 25 0 0 0 0 0 25 1 20 99 26 0 100 96 93 0
Pseudomonas luteola 86 75 0 0 94 0 0 0 0 25 13 84 0 1 0 1 1 15 1 85 0 30 0 100 91 94 0
Pseudomonas oryzihabitans 0 0 0 0 89 0 0 0 0 25 1 10 0 1 0 1 0 10 0 45 0 7 0 100 99 99 0
Non-fermenter spp 1 1 0 0 37 0 1 0 0 15 9 9 0 0 0 1 1 1 1 1 93 48 35 99 85 49 0
Shewanella putrefaciens group 0 0 0 80 75 75 1 0 0 0 75 1 0 0 0 0 1 0 0 2 99 96 0 100 96 9 0
Stenotrophomonas maltophilia 70 0 75 1 75 1 0 0 0 0 90 1 0 0 0 0 0 0 0 0 1 26 1 100 91 49 0
* Brucella spp possible / möglich / posible / possibile / possível / / möjlig / mulig / Mo liwo .
bioMérieux SA III
api® 20 E TM 07584J - xl - 2010/05
bioMérieux SA IV
api® 20 E TM 07584J - xl - 2010/05
bioMérieux SA V
api® 20 E TM 07584J - xl - 2010/05
(1) Une très légère couleur jaune est également positive / A very pale yellow should also be considered positive / Auch eine nur ganz leichte Gelbfärbung ist als positiv zu bewerten / Un color amarillo muy
ligero también implica resultado positivo / Una leggerissima colorazione gialla è comunque positiva / Uma cor amarela muito ligeira é também positiva. /
/ En mycket ljust gul färgning ska också anses som positiv / En meget lys gul skal også betragtes som positiv / Nawet bardzo blady ó ty kolor nale y rozpatrywa jako pozytywny.
(2) Une couleur orange apparaissant après 36-48 H d'incubation doit être considérée négative / An orange color after 36-48 hours incubation must be considered negative / Eine orange Verfärbung nach
einer 36-48-stündigen Inkubation wird als negativ bewertet / La aparición de un color naranja tras 36-48 H de incubación debe considerarse negativa / Se dopo 36-48 ore di incubazione appare una
colorazione arancione, la reazione deve essere considerata negativa / Uma cor laranja após 36-48 H de incubação deve ser considerada negativa. / 36-48
/ En orange färg efter 36-48 timmars inkubation ska anses negativ / En orange farve efter 36-48 timers inkubation skal betragtes som negativ / Pomara czowy
kolor po 36-48 godzinach inkubacji nale y uwa a za negatywny.
(3) Lecture dans la cupule (zone aérobie) / Reading made in the cupule (aerobic) / Ablesung im Becher (aerober Bereich) / Lectura en la cúpula (zona aerobia) / Lettura nella cupola (zona aerobia) / Leitura
na cúpula (zona aeróbia). / ( ) / Avläsning utförd i kupolen (aerob) / Aflæsning foretaget i brønden (aerob) / Odczytu dokona we wg bieniu (warunki tlenowe).
(4) La fermentation commence dans la partie inférieure des tubes, l'oxydation commence dans la cupule / Fermentation begins in the lower portion of the tubes, oxidation begins in the cupule / Die
Fermentation beginnt im unteren Teil der Röhrchens, die Oxidation im Becher / La fermentación comienza en la parte inferior de los tubos, mientras que la oxidación empieza en la cúpula / La
fermentazione comincia nella parte inferiore delle microprovette, mentre l'ossidazione comincia nella cupola / A fermentação começa na parte inferior dos tubos, a oxidação começa na cúpula. /
, / Jäsning börjar i brunnens nedre delar, oxidation börjar i kupolen / Fermentation starter i den nederste del af
rørene, oxidation starter i brønden / Fermentacja zachodzi w najni szej cz ci probówki, utlenianie we wg bieniu.
(5) Une légère coloration rose apparaissant après 10 minutes doit être lue négative / A slightly pink color after 10 minutes should be considered negative / Eine nach 10 min auftretende schwache rosa
Verfärbung wird als negativ bewertet / Una ligera coloración rosa, que aparece tras 10 minutos, debe ser leída como negativa / Una debole colorazione rosa che appaia dopo oltre 10 minuti deve
essere considerata negativa / Uma ligeira coloração rosa depois de 10 minutos deve ser considerada negativa. / 10 / En
svagt rosa färg efter 10 minuter ska anses negativ / En let lyserød farve efter 10 minutter skal betragtes som negativ / S abo ró owy kolor po 10 minutach nale y uwa a za negatywny.
bioMérieux SA VI
api® 20 E TM
Les quantités indiquées peuvent être ajustées en fonction des titres des matières premières / The quantities indicated may be adjusted depending on the titer of the raw materials used / Die
angegebenen Mengen können je nach Konzentration der verwendeten Ausgangsmaterialien angeglichen werden. / Las cantidades indicadas pueden ser ajustadas en función de los títulos de las
materias primas / Le quantità indicate possono essere aggiustate in funzione dei titoli delle materie prime / As quantidades indicadas podem ser ajustadas em função dos títulos das matérias-primas./
/ Den angivna mängden kan justeras beroende på titern av de använda råmaterialen / De
angivne mængder kan justeres, afhængigt af titeren for de anvendte råmaterialer / Wskazane st enia mog by regulowane w zale no ci od miana u ytego surowego materia u.
Certaines cupules contiennent des composants d’origine animale, notamment des peptones / Certain cupules contain products of animal origin, notably peptones / Einige Näpfchen enthalten
Bestandteile tierischen Ursprungs, vor allem Peptone / Ciertas cúpulas contienen componentes de origen animal, en concreto peptonas / Alcune cupole contengono dei componenti di origine animale,
in particolare dei peptoni / Algumas cúpulas contêm componentes de origem animal, nomeadamente, peptonas. / , / Vissa
kupoler innehåller produkter av animaliskt ursprung, i synnerhet peptoner / Visse brønde indeholder produkter af animalsk oprindelse, specielt peptoner / Niektóre mikroprobówki zawieraj produkty
pochodzenia zwierz cego,zw aszcza peptony.
bioMérieux SA VII
api® 20 E TM 07584J - xl - 2010/05
bioMérieux SA VIII
api® 20 E TM 07584J - xl - 2010/05
Se handhavandebeskrivningen / Se brugsanvisning
Sprawd w instrukcji obs ugi
Contenu suffisant pour "n" tests
Contains sufficient for <n> tests
Inhalt ausreichend für <n> Prüfungen
Contenido suficiente para <n> ensayos
Contenuto sufficiente per "n" saggi
Conteúdo suficiente para “n” ensaios
« »
Räcker till "n" antal tester
Indeholder tilstrækkeligt til "n" test
Wystarczy na wykonanie <n> testów
®
20 E TM IVD
Sistema de identificação das Enterobacteriaceae e outros bacilos Gram negativos não fastidiosos
bioMérieux SA Português - 1
api® 20 E TM 07584J - pt - 2010/05
bioMérieux SA Português - 2
api® 20 E TM 07584J - pt - 2010/05
Nalguns casos, pode haver necessidade de efectuar - Mobilidade (MOB) : Inocular uma ampola de API M
testes complementares por o perfil de 7 algarismos não Medium (cfr folheto informativo).
ser iuficientemente discriminativo: - Cultura em gelose MacConkey (McC): Semear um meio
- Redução dos nitratos em nitritos (NO2) e em azoto (N2) : de Mac Conkey (cfr folheto informativo).
adicionar 1 gota dos reagentes NIT 1 e NIT 2 no tubo - Oxidação da glucose (OF-O) : Inocular uma ampola de
GLU. Esperar 2 a 5 minutos. Uma cor vermelha indica API OF Medium (cfr folheto informativo).
uma reacção positiva (NO2). Uma reacção negativa - Fermentação da glucose (OF-F) : Inocular uma ampola
(coloração amarela) pode ser devida à produção de de API OF Medium (cfr folheto informativo).
azoto (eventualmente assinalada pela presença de Estes testes complementares, mencionados na
bolhas mínimas): adicionar 2 a 3 mg de reagente Zn na introdução (Código dos perfis) do Catálogo Analítico,
cúpula GLU. Após 5 minutos, se um tubo permanecer podem ser utilizados para constituir um perfil de
amarelo indica uma reacção positiva (N2) a anotar na 9 algarismos, identificável com o programa de
ficha de resultados. Se a cúpula estiver laranja- identificação.
vermelha, a reacção é negativa, os nitratos ainda
presentes no tubo foram reduzidos a nitritos pelo Zinco.
Esta reacção é especialmente interessante para os
bacilos Gram negativos oxidase positiva.
5 315 173 (57) Enterobacter gergoviae
NOTA: Pelas mesmas razões que as do teste indol
(consultar a nota do parágrafo "Leitura da galeria"), o Podem ser propostos testes suplementares se houver
teste de redução dos nitratos deve efectuar-se por último. fraca discriminação. Consultar o programa ou o Catálogo
Analítico.
CONTROLO DE QUALIDADE
Os meios, galerias e reagentes são objecto de controlos de qualidade sistemáticos nas diferentes etapas do seu fabrico.
O Controlo de Qualidade Mínimo pode ser utilizado para verificar que as condições de armazenamento e transporte
não têm impacto no comportamento funcional da galeria API 20 E. Este controlo pode ser efectuado em conformidade
®
com as instruções e critérios esperados acima em relação ao referencial CLSI M50-A Quality Control for Commercial
Microbial Identification Systems.
®
O controlo pode ser efectuado utilizando a estirpe/cepa Proteus mirabilis ATCC 35659 para avaliar o comportamento
funcional dos testes ODC e ARA. Os estudos efectuados pela bioMérieux demonstraram que na galeria API 20 E, os
testes ODC e ARA são os testes mais sensíveis. Aquando do controlo, a integridade da galeria pode ser verificada
utilizando a estirpe/cepa Proteus mirabilis ATCC 35659.
No caso se ser exigido um Controlo de Qualidade Completo para esta galeria, as cinco estirpes/cepas seguintes
deverão ser testadas para verificar as reacções positivas e negativas da maioria dos testes da galeria API 20 E.
1 Proteus mirabilis ATCC 35659 4. Escherichia coli ATCC 25922
2. Stenotrophomonas maltophilia ATCC 51331 5. Klebsiella pneumoniae ssp pneumoniae ATCC 35657
3. Enterobacter cloacae ATCC 13047
ATCC : American Type Culture Collection, 10801 University Boulevard, Manassas, VA 20110-2209, USA.
ONPG ADH LDC ODC CIT H2S URE TDA IND VP GEL GLU MAN INO SOR RHA SAC MEL AMY ARA NO2 N2*
1. – – – + V + + + – – V + – – – – V – – – + –
2. + – V – V – – – – – + – – – – – – – – – – –
3. + + – V + – – – – + – + + V + + + + + + + –
4. + – + + – – – – + – – + + – + + – + – + + –
5. + – + – + – V – – V – + + + + + + + + + + –
* O estado N2 (+) pode ser observado para as estirpes/cepas ATCC 13047, ATCC 25922 e ATCC 35657.
Perfil obtido após 24-48 H de incubação para a estirpe/cepa ATCC 51331, a partir de colónias cultivadas em gelose Trypcase Soja +
sangue.
Perfis obtidos após 18-24 H de incubação para as outras estirpes/cepas, a partir de colónias cultivadas em gelose Trypcase Soja +
sangue.
Suspensão bacteriana preparada em API NaCl 0,85 % Medium.
É da responsabilidade do utilizador assegurar que o controlo de qualidade é efectuado em conformidade com a
legislação local em vigor.
bioMérieux SA Português - 3
api® 20 E TM 07584J - pt - 2010/05
No caso de identificação de Salmonella ou Shigella, Outros bacilos Gram negativos não fastidiosos:
deve efectuar-se uma identificação serológica para Foram testadas 2386 estirpes/cepas de diversas
confirmar a identificação bacteriana. origens e estirpes/cepas de colecção perctencentes
Os bacilos Gram negativos não fermentadores, às espécies da base de dados:
isolados de doentes com mucoviscidose, podem criar - 90,32 % das estirpes/cepas foram correctamente
perfis bioquímicos atípicos susceptíveis de alterar a identificadas (com ou sem testes complementares).
sua identificação. - 6,16 % das estirpes/cepas não foram identificadas.
Devem apenas ser utilizadas as culturas puras que - 3,52 % das estirpes/cepas foram mal identificadas.
contenham um único tipo de microrganismo.
ELIMINAÇÃO DE RESÍDUOS
RESULTADOS ESPERADOS Eliminar os reagentes utilizados ou não utilizados bem
Consultar o Quadro de Identificação no final deste como os materiais descartáveis contaminados em
folheto informativo para saber os resultados esperados conformidade com os procedimentos relativos aos
para as diferentes reacções bioquímicas. produtos infecciosos ou potencialmente infecciosos.
É da responsabilidade de cada laboratório gerir os
COMPORTAMENTO FUNCIONAL resíduos e os efluentes que este produz consoante a
Enterobacteriaceae : sua natureza e o seu perigo, e assegurar (ou fazer
Foram testadas 5514 estirpes/cepas de diversas assegurar) o tratamento e a eliminação em
origens e estirpes/cepas de colecção pertencentes às conformidade com as regulamentações aplicáveis.
espécies da base de dados:
- 92,80 % das estirpes/cepas foram correctamente
identificadas (com ou sem testes complementares).
- 4,61 % das estirpes/cepas não foram identificadas.
- 2,59 % das estirpes/cepas foram mal identificadas.
PROCEDIMENTO p. I
QUADRO DE IDENTIFICAÇÃO p. II
QUADRO DE LEITURA p. IV
TESTES COMPLEMENTARES p. VII
BIBLIOGRAFIA p. VIII
QUADRO DOS SÍMBOLOS p. IX
A BIOMERIEUX, o logotipo azul, API e apiweb são marcas utilizadas, depositadas e/ou registadas, propriedade exclusiva da bioMérieux SA ou de uma
das suas filiais.
CLSI é uma marca, propriedade exclusiva da Clinical and Laboratory Standards Institute, Inc.
ATCC é uma marca da propriedade exclusiva da American Type Culture Collection.
As outras marcas e nomes de produtos mencionados neste documento são marcas comerciais dos respectivos proprietários.
Brasil: Distribuído por bioMérieux Brasil, S.A. - Estrada do Mapuá, 491 - Jacarepaguá - R.J. - CEP 22710-261
CNPJ: 33.040.635/0001-71
Atendimento ao Consumidor Tel.: 0800-264848
Prazo de Validade, N° de Lote, N° de Registro de Ministério da Saúde e Responsável Técnico:
VIDE EMBALAGEM
OX
- | CIT |
- | VP |
- | GEL |
API 20 E
ADH ODC
H2S - URE
36°C
18:00-24:00 / 48:00 ±
2°C
Tests < 3
(y compris / including /
einschließlich / incluído /
compreso / incluindo / API 20 E
/ inklusive /
inklusiv / w czaj c test GLU) TDA : TDA
IND : JAMES
VP : VP 1 + VP 2
GLU (NO2) : NIT 1 + NIT 2 (+Zn)
API 20 E
+ - + - + -
bioMérieux SA I
api® 20 E TM 07584J - xl - 2010/05
TABLEAU D'IDENTIFICATION / IDENTIFICATION TABLE / PROZENTTABELLE / TABLA DE IDENTIFICACION / TABELLA DI IDENTIFICAZIONE / QUADRO DE IDENTIFICAÇÃO
/ IDENTIFIKATIONSTABEL / IDENTIFIERINGSTABELL / TABELA IDENTYFIKACYJNA
% de réactions positives après 18-24 / 48 h à 36°C ± 2°C / % of positive reactions after 18-24 / 48 hrs. at 36°C ± 2°C / % der positiven Reaktionen nach 18-24 / 48 h bei 36°C ± 2°C /
% de las reacciones positivas después de 18-24 / 48 H a 36°C ± 2°C / % di reazioni positive dopo 18-24 / 48 ore a 36°C ± 2°C / % de reacções positivas após 18-24 / 48 h a 36º C ± 2º C /
% 18-24 / 48 36°C ± 2°C / % positiva reaktioner efter 18-24 / 48 timmar vid 36°C ± 2°C / % af positive reaktioner efter 18-24 / 48 timer ved 36°C ± 2°C /
% pozytywnych reakcji po 18-24 / 48 godzinach w 36°C ± 2°C
API 20 E V4.1 ONPG ADH LDC ODC CIT H2S URE TDA IND VP GEL GLU MAN INO SOR RHA SAC MEL AMY ARA OX NO2 N2 MOB McC OF/O OF/F
Buttiauxella agrestis 100 0 0 85 25 0 0 0 0 0 0 100 100 0 1 99 0 92 99 100 0 100 0 100 100 100 100
Cedecea davisae 99 89 0 99 75 0 0 0 0 89 0 100 100 10 0 0 100 0 100 1 0 99 0 87 100 100 100
Cedecea lapagei 99 99 0 0 75 0 0 0 0 90 0 100 99 0 0 0 0 1 100 1 0 99 0 87 100 100 100
Citrobacter braakii 50 45 0 99 75 81 1 0 4 0 0 100 100 1 100 100 1 91 99 99 0 100 0 95 100 100 100
Citrobacter freundii 90 24 0 0 75 75 1 0 1 0 0 100 99 25 99 99 99 82 40 99 0 98 0 95 100 100 100
Citrobacter koseri/amalonaticus 99 75 0 100 97 0 1 0 99 0 0 100 100 25 99 99 1 1 98 99 0 100 0 95 100 100 100
Citrobacter koseri/farmeri 99 2 0 100 25 0 1 0 99 0 0 100 100 1 99 99 99 80 99 99 0 100 0 95 100 100 100
Citrobacter youngae 100 50 0 1 80 80 0 0 1 0 0 100 100 0 95 100 1 0 25 100 0 85 0 95 100 100 100
Edwardsiella hoshinae 0 0 100 99 50 94 0 0 99 0 0 100 100 0 0 1 100 0 0 1 0 100 0 100 100 100 100
Edwardsiella tarda 0 0 100 99 1 75 0 0 99 0 0 100 0 0 0 0 0 0 0 0 0 100 0 98 100 100 100
Enterobacter aerogenes 99 0 99 98 82 0 1 0 0 85 0 99 99 99 99 99 99 99 99 99 0 100 0 97 100 100 100
Enterobacter amnigenus 1 99 25 0 99 40 0 0 0 0 75 0 100 100 0 1 100 99 99 99 99 0 100 0 92 100 100 100
Enterobacter amnigenus 2 99 80 0 99 80 0 0 0 0 75 0 100 100 0 99 100 1 99 99 99 0 100 0 100 100 100 100
Enterobacter asburiae 100 25 0 99 80 0 0 0 0 10 0 100 99 25 100 0 99 0 100 100 0 100 0 95 100 100 100
Enterobacter cancerogenus 100 75 0 99 99 0 0 0 0 89 0 100 100 0 1 100 1 1 100 100 0 100 0 99 100 100 100
Enterobacter cloacae 98 82 1 92 90 0 1 0 0 85 0 99 99 12 90 85 96 90 99 99 0 100 0 95 100 100 100
Enterobacter gergoviae 99 0 32 100 75 0 99 0 0 90 0 100 99 23 1 100 99 100 99 100 0 100 0 90 100 100 100
Enterobacter intermedius 99 0 0 99 1 0 0 0 0 2 0 100 97 0 88 99 40 100 99 99 0 100 0 92 100 100 100
Enterobacter sakazakii 100 96 0 91 94 0 1 0 25 91 10 100 100 75 8 99 99 99 99 99 0 100 0 96 100 100 100
Escherichia coli 1 90 1 74 70 0 1 3 0 89 0 0 99 98 1 91 82 36 75 3 99 0 100 0 95 100 100 100
Escherichia coli 2 26 1 45 20 0 1 1 0 50 0 0 99 90 1 42 30 3 3 1 70 0 98 0 5 100 100 100
Escherichia fergusonii 96 1 99 100 1 0 0 0 99 0 0 100 99 1 0 87 0 1 99 99 0 100 0 93 100 100 100
Escherichia hermannii 100 0 1 100 1 0 0 0 99 0 0 100 100 0 0 99 25 0 99 99 0 100 0 99 100 100 100
Escherichia vulneris 100 30 50 0 0 0 0 0 0 0 0 100 100 0 1 95 7 95 95 99 0 100 0 100 100 100 100
Ewingella americana 98 0 0 0 75 0 0 0 0 95 1 99 99 0 0 1 0 1 50 1 0 100 0 60 100 100 100
Hafnia alvei 1 75 0 99 98 50 0 10 0 0 50 0 99 99 0 1 99 0 0 25 99 0 100 0 85 100 100 100
Hafnia alvei 2 50 0 99 99 1 0 1 0 0 10 0 99 98 0 1 1 1 0 0 1 0 100 0 0 100 100 100
Klebsiella oxytoca 99 0 80 0 89 0 78 0 99 80 0 100 100 99 100 99 99 100 100 100 0 100 0 0 100 100 100
Klebsiella pneumoniae ssp ozaenae 94 18 25 1 18 0 1 0 0 1 0 99 96 57 66 58 20 80 97 85 0 92 0 0 100 100 100
Klebsiella pneumoniae ssp pneumoniae 99 0 73 0 86 0 75 0 0 90 0 100 99 99 99 99 99 99 99 99 0 100 0 0 100 100 100
Klebsiella pneumoniae ssp rhinoscleromatis 1 0 0 0 0 0 0 0 0 0 0 99 100 90 90 75 75 1 99 10 0 100 0 0 100 100 100
Kluyvera spp 95 0 25 99 60 0 0 0 80 0 0 100 99 0 25 93 89 99 99 99 0 95 0 94 100 100 100
Leclercia adecarboxylata 99 0 0 0 0 0 1 0 99 0 1 100 99 0 2 100 66 99 99 100 0 100 0 100 100 100 100
Moellerella wisconsensis 97 0 0 0 40 0 0 0 15 1 0 100 1 0 0 0 100 99 0 0 0 90 0 0 100 100 100
Morganella morganii 1 0 10 98 1 1 99 93 99 0 0 99 0 0 0 0 1 0 0 0 0 88 0 95 100 100 100
Pantoea spp 1 85 1 0 0 13 0 1 0 1 9 1 100 99 1 26 1 98 26 59 61 0 85 0 85 100 100 100
Pantoea spp 2 99 1 0 0 99 0 1 0 53 62 4 100 99 36 82 90 98 81 99 99 0 85 0 85 100 100 100
Pantoea spp 3 99 1 0 0 21 0 1 0 1 86 15 100 99 34 1 97 93 23 65 97 0 85 0 85 100 100 100
Pantoea spp 4 86 1 0 0 29 0 1 0 59 1 1 99 100 10 32 99 72 89 99 99 0 85 0 85 100 100 100
Proteus mirabilis 1 0 0 99 50 75 99 98 1 1 82 98 0 0 0 0 1 0 0 0 0 93 0 95 100 100 100
Proteus penneri 1 0 0 0 1 20 100 99 0 0 50 99 0 0 0 0 100 0 1 0 0 99 0 85 100 100 100
Proteus vulgaris group 1 0 0 0 12 83 99 99 92 0 74 99 1 1 0 1 89 0 66 1 0 100 0 94 100 100 100
Providencia alcalifaciens/rustigianii 0 0 0 0 80 0 0 100 99 0 0 99 1 1 0 0 1 0 0 1 0 100 0 96 100 100 100
Providencia rettgeri 1 1 0 0 74 0 99 99 90 0 0 98 82 78 1 50 25 0 40 1 0 98 0 94 100 100 100
Providencia stuartii 1 0 0 0 85 0 30 98 95 0 0 98 3 80 0 0 15 0 0 0 0 100 0 85 100 100 100
Rahnella aquatilis 100 0 0 0 50 0 0 1 0 99 0 100 100 0 98 99 100 97 100 98 0 100 0 6 100 100 100
Raoultella ornithinolytica 100 0 99 99 99 0 85 0 100 65 0 100 100 99 100 100 100 100 100 100 0 100 0 0 100 100 100
Raoultella terrigena 100 0 99 6 52 0 0 0 0 75 0 99 99 99 99 99 100 100 100 99 0 100 0 0 100 100 100
Salmonella choleraesuis ssp arizonae 98 75 97 98 75 99 0 0 1 0 0 100 99 0 99 99 1 78 0 99 0 100 0 99 100 100 100
Salmonella choleraesuis ssp choleraesuis 0 15 99 99 6 64 0 0 0 0 0 100 99 0 98 99 0 20 0 0 0 100 0 95 100 100 100
Salmonella ser.Gallinarum 0 1 100 1 0 25 0 0 0 0 0 100 100 0 0 1 0 0 0 100 0 100 0 0 100 100 100
bioMérieux SA II
api® 20 E TM 07584J - xl - 2010/05
API 20 E V4.1 ONPG ADH LDC ODC CIT H2S URE TDA IND VP GEL GLU MAN INO SOR RHA SAC MEL AMY ARA OX NO2 N2 MOB McC OF/O OF/F
Salmonella ser.Paratyphi A 0 5 0 99 0 1 0 0 0 0 0 100 99 0 99 98 0 96 0 99 0 100 0 95 100 100 100
Salmonella ser.Pullorum 0 1 75 100 0 85 0 0 0 0 0 100 100 0 0 100 0 0 0 75 0 100 0 0 100 100 100
Salmonella typhi 0 1 99 0 0 8 0 0 0 0 0 100 99 0 99 0 0 99 0 0 0 100 0 97 100 100 100
Salmonella spp 1 56 82 93 65 83 0 0 1 0 1 99 100 40 99 86 1 90 1 99 1 100 0 95 100 100 100
Serratia ficaria 99 0 0 0 100 0 0 0 0 40 90 100 100 50 99 74 99 99 100 99 0 92 0 100 100 100 100
Serratia fonticola 99 0 73 99 75 0 0 0 0 0 0 100 100 97 100 99 30 99 99 99 0 99 0 91 100 100 100
Serratia liquefaciens 95 1 78 98 80 0 2 0 0 59 65 100 99 80 98 2 99 72 97 97 0 100 0 95 100 100 100
Serratia marcescens 94 0 95 95 96 0 25 0 1 70 87 100 99 85 98 1 99 68 97 25 0 95 0 97 100 100 100
Serratia odorifera 1 95 0 95 99 95 0 0 0 99 50 99 100 99 99 99 99 99 99 99 99 0 99 0 100 100 100 100
Serratia odorifera 2 95 0 96 1 95 0 0 0 99 50 99 100 99 99 99 99 1 99 99 95 0 99 0 100 100 100 100
Serratia plymuthica 99 0 0 0 65 0 0 0 0 65 50 100 90 70 70 1 99 85 98 98 0 99 0 50 100 100 100
Serratia rubidaea 99 0 30 0 92 0 1 0 0 71 82 99 99 75 1 3 99 95 99 99 0 100 0 85 100 100 100
Shigella spp 1 0 0 1 0 0 0 0 29 0 0 99 63 0 7 7 1 20 0 50 0 100 0 0 100 100 100
Shigella sonnei 96 0 0 93 0 0 0 0 0 0 0 99 99 0 1 75 1 1 0 99 0 100 0 0 100 100 100
Yersinia enterocolitica 80 0 0 90 0 0 98 0 50 5 0 99 99 25 98 1 99 4 75 75 0 98 0 2 100 100 100
Yersinia frederiksenii/intermedia 99 0 0 75 1 0 99 0 99 1 0 100 99 25 99 99 99 1 99 99 0 98 0 5 100 100 100
Yersinia kristensenii 80 0 0 80 0 0 99 0 97 0 0 100 99 10 99 0 0 0 99 99 0 98 0 5 100 100 100
Yersinia pestis 68 0 0 0 0 0 0 0 0 1 0 99 99 0 70 0 0 0 30 30 0 47 0 0 99 100 100
Yersinia pseudotuberculosis 98 0 0 0 1 0 99 0 0 0 0 99 97 0 0 75 0 50 25 50 0 95 0 0 100 100 100
Aeromonas hydrophila gr. 1 98 90 25 1 25 0 0 0 85 25 90 99 99 1 3 5 97 1 75 75 100 97 0 95 99 99 99
Aeromonas hydrophila gr. 2 99 97 80 1 80 0 0 0 85 80 97 97 99 9 9 1 80 1 75 5 100 97 0 95 99 99 99
Aeromonas salmonicida ssp salmonicida 1 60 1 0 0 0 0 0 1 0 75 50 54 0 0 0 0 0 1 0 100 98 0 1 99 99 99
Grimontia hollisae 1 0 0 0 0 0 0 0 94 0 0 10 0 0 0 0 0 0 0 0 100 100 0 0 99 99 99
Photobacterium damselae 1 99 75 0 1 0 98 0 0 10 1 50 0 0 0 0 1 0 0 0 100 100 0 25 99 99 99
Plesiomonas shigelloides 95 99 100 100 0 0 0 0 100 0 0 99 0 99 0 0 0 0 0 0 100 99 0 95 99 99 99
Vibrio alginolyticus 0 0 98 75 60 0 1 0 100 10 75 99 100 0 1 0 100 0 10 1 100 47 0 100 99 94 94
Vibrio cholerae 98 1 94 97 75 0 0 0 99 58 92 98 98 0 0 0 94 0 5 0 100 96 0 100 96 99 99
Vibrio fluvialis 95 99 0 0 1 0 0 0 80 0 75 75 80 0 1 0 75 0 36 75 100 100 0 100 99 99 99
Vibrio mimicus 99 0 99 99 50 0 0 0 99 1 99 99 99 0 0 0 0 0 0 0 100 95 0 100 95 99 99
Vibrio parahaemolyticus 0 0 100 99 50 0 1 0 100 1 75 100 99 0 0 1 1 0 12 50 100 63 0 100 98 99 99
Vibrio vulnificus 99 0 91 90 25 0 0 0 99 1 99 99 75 0 0 0 1 0 90 0 99 54 0 100 99 99 99
Pasteurella aerogenes 99 0 0 80 0 0 99 0 0 0 0 99 0 97 0 1 99 0 0 75 75 100 0 0 100 100 100
Pasteurella multocida 1 4 0 0 25 0 0 0 0 99 0 0 29 1 0 1 0 75 0 0 0 99 90 0 0 2 23 23
Pasteurella multocida 2 7 0 0 45 0 0 0 0 99 0 0 44 99 0 99 0 99 0 0 0 89 90 0 0 2 23 23
Pasteurella pneumotropica/ Mannheimia haemolytica 60 0 1 10 0 0 25 0 15 7 3 35 12 12 12 1 35 1 2 1 80 99 0 0 9 33 33
Acinetobacter baumannii/calcoaceticus 0 0 0 0 51 0 1 0 0 5 5 99 0 0 0 0 0 99 1 99 0 3 0 0 90 98 0
Bordetella/Alcaligenes/Moraxella spp * 0 0 0 0 52 0 14 1 0 25 1 0 0 0 0 0 0 0 0 0 95 62 1 75 75 0 0
Burkholderia cepacia 50 0 25 16 78 0 0 0 0 1 43 60 1 0 0 0 13 0 7 20 90 40 0 99 88 97 0
Chromobacterium violaceum 0 99 0 0 75 0 0 0 14 0 99 99 0 0 0 0 10 0 0 0 99 75 0 99 99 99 99
Chryseobacterium indologenes 5 0 0 0 12 0 90 0 75 0 80 0 0 0 0 0 0 0 0 0 99 20 0 0 57 90 10
Chryseobacterium meningosepticum 77 0 0 0 20 0 1 0 85 0 90 0 0 0 0 0 0 0 0 0 99 6 0 0 48 93 6
Eikenella corrodens 0 0 75 99 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 100 95 0 1 1 49 49
Myroides /Chryseobacterium indologenes 0 0 0 0 50 0 75 0 0 1 75 0 0 0 0 0 0 0 0 0 99 0 0 0 84 2 2
Ochrobactrum anthropi 15 0 0 0 30 0 25 1 0 15 0 1 0 0 0 0 0 0 0 10 90 42 60 99 99 47 0
Pseudomonas aeruginosa 0 89 0 0 92 0 25 0 0 1 75 50 0 0 0 0 1 10 1 25 97 12 56 97 100 98 0
Pseudomonas fluorescens/putida 0 75 0 0 75 0 0 0 0 10 27 25 0 0 0 0 0 25 1 20 99 26 0 100 96 93 0
Pseudomonas luteola 86 75 0 0 94 0 0 0 0 25 13 84 0 1 0 1 1 15 1 85 0 30 0 100 91 94 0
Pseudomonas oryzihabitans 0 0 0 0 89 0 0 0 0 25 1 10 0 1 0 1 0 10 0 45 0 7 0 100 99 99 0
Non-fermenter spp 1 1 0 0 37 0 1 0 0 15 9 9 0 0 0 1 1 1 1 1 93 48 35 99 85 49 0
Shewanella putrefaciens group 0 0 0 80 75 75 1 0 0 0 75 1 0 0 0 0 1 0 0 2 99 96 0 100 96 9 0
Stenotrophomonas maltophilia 70 0 75 1 75 1 0 0 0 0 90 1 0 0 0 0 0 0 0 0 1 26 1 100 91 49 0
* Brucella spp possible / möglich / posible / possibile / possível / / möjlig / mulig / Mo liwo .
bioMérieux SA III
api® 20 E TM 07584J - xl - 2010/05
bioMérieux SA IV
api® 20 E TM 07584J - xl - 2010/05
bioMérieux SA V
api® 20 E TM 07584J - xl - 2010/05
(1) Une très légère couleur jaune est également positive / A very pale yellow should also be considered positive / Auch eine nur ganz leichte Gelbfärbung ist als positiv zu bewerten / Un color amarillo muy
ligero también implica resultado positivo / Una leggerissima colorazione gialla è comunque positiva / Uma cor amarela muito ligeira é também positiva. /
/ En mycket ljust gul färgning ska också anses som positiv / En meget lys gul skal også betragtes som positiv / Nawet bardzo blady ó ty kolor nale y rozpatrywa jako pozytywny.
(2) Une couleur orange apparaissant après 36-48 H d'incubation doit être considérée négative / An orange color after 36-48 hours incubation must be considered negative / Eine orange Verfärbung nach
einer 36-48-stündigen Inkubation wird als negativ bewertet / La aparición de un color naranja tras 36-48 H de incubación debe considerarse negativa / Se dopo 36-48 ore di incubazione appare una
colorazione arancione, la reazione deve essere considerata negativa / Uma cor laranja após 36-48 H de incubação deve ser considerada negativa. / 36-48
/ En orange färg efter 36-48 timmars inkubation ska anses negativ / En orange farve efter 36-48 timers inkubation skal betragtes som negativ / Pomara czowy
kolor po 36-48 godzinach inkubacji nale y uwa a za negatywny.
(3) Lecture dans la cupule (zone aérobie) / Reading made in the cupule (aerobic) / Ablesung im Becher (aerober Bereich) / Lectura en la cúpula (zona aerobia) / Lettura nella cupola (zona aerobia) / Leitura
na cúpula (zona aeróbia). / ( ) / Avläsning utförd i kupolen (aerob) / Aflæsning foretaget i brønden (aerob) / Odczytu dokona we wg bieniu (warunki tlenowe).
(4) La fermentation commence dans la partie inférieure des tubes, l'oxydation commence dans la cupule / Fermentation begins in the lower portion of the tubes, oxidation begins in the cupule / Die
Fermentation beginnt im unteren Teil der Röhrchens, die Oxidation im Becher / La fermentación comienza en la parte inferior de los tubos, mientras que la oxidación empieza en la cúpula / La
fermentazione comincia nella parte inferiore delle microprovette, mentre l'ossidazione comincia nella cupola / A fermentação começa na parte inferior dos tubos, a oxidação começa na cúpula. /
, / Jäsning börjar i brunnens nedre delar, oxidation börjar i kupolen / Fermentation starter i den nederste del af
rørene, oxidation starter i brønden / Fermentacja zachodzi w najni szej cz ci probówki, utlenianie we wg bieniu.
(5) Une légère coloration rose apparaissant après 10 minutes doit être lue négative / A slightly pink color after 10 minutes should be considered negative / Eine nach 10 min auftretende schwache rosa
Verfärbung wird als negativ bewertet / Una ligera coloración rosa, que aparece tras 10 minutos, debe ser leída como negativa / Una debole colorazione rosa che appaia dopo oltre 10 minuti deve
essere considerata negativa / Uma ligeira coloração rosa depois de 10 minutos deve ser considerada negativa. / 10 / En
svagt rosa färg efter 10 minuter ska anses negativ / En let lyserød farve efter 10 minutter skal betragtes som negativ / S abo ró owy kolor po 10 minutach nale y uwa a za negatywny.
bioMérieux SA VI
api® 20 E TM
Les quantités indiquées peuvent être ajustées en fonction des titres des matières premières / The quantities indicated may be adjusted depending on the titer of the raw materials used / Die
angegebenen Mengen können je nach Konzentration der verwendeten Ausgangsmaterialien angeglichen werden. / Las cantidades indicadas pueden ser ajustadas en función de los títulos de las
materias primas / Le quantità indicate possono essere aggiustate in funzione dei titoli delle materie prime / As quantidades indicadas podem ser ajustadas em função dos títulos das matérias-primas./
/ Den angivna mängden kan justeras beroende på titern av de använda råmaterialen / De
angivne mængder kan justeres, afhængigt af titeren for de anvendte råmaterialer / Wskazane st enia mog by regulowane w zale no ci od miana u ytego surowego materia u.
Certaines cupules contiennent des composants d’origine animale, notamment des peptones / Certain cupules contain products of animal origin, notably peptones / Einige Näpfchen enthalten
Bestandteile tierischen Ursprungs, vor allem Peptone / Ciertas cúpulas contienen componentes de origen animal, en concreto peptonas / Alcune cupole contengono dei componenti di origine animale,
in particolare dei peptoni / Algumas cúpulas contêm componentes de origem animal, nomeadamente, peptonas. / , / Vissa
kupoler innehåller produkter av animaliskt ursprung, i synnerhet peptoner / Visse brønde indeholder produkter af animalsk oprindelse, specielt peptoner / Niektóre mikroprobówki zawieraj produkty
pochodzenia zwierz cego,zw aszcza peptony.
bioMérieux SA VII
api® 20 E TM 07584J - xl - 2010/05
bioMérieux SA VIII
api® 20 E TM 07584J - xl - 2010/05
Se handhavandebeskrivningen / Se brugsanvisning
Sprawd w instrukcji obs ugi
Contenu suffisant pour "n" tests
Contains sufficient for <n> tests
Inhalt ausreichend für <n> Prüfungen
Contenido suficiente para <n> ensayos
Contenuto sufficiente per "n" saggi
Conteúdo suficiente para “n” ensaios
« »
Räcker till "n" antal tester
Indeholder tilstrækkeligt til "n" test
Wystarczy na wykonanie <n> testów
PW4
ISOLATION AND IDENTIFICATION OF LACTIC ACID BACTERIA
Master in Microbiology
1. Isolation
1.1 For each supplied sample prepare, under aseptic conditions, a 10% suspension (% w/v
or % v/v depending on the type of sample) in 10 ml of saline solution (0.9% NaCl) or
peptonated water (0.1% peptone).
If necessary, fragment, grind or macerate the sample in advance for homogenisation.
1.2 After vortexing, plaque an aliquot of each sample on a MRS medium plate pH 6.5
(1.5% agar) and/or pH 5.5 (2% agar) containing 0.01 % cycloheximide.
To obtain isolated colonies, dilute previously (10-2) and plate 0.1 ml or alternatively
inoculate a loop of the original suspension.
1.4 When checking for colonies, microscopic examination should be caried out to detect the
presence of gram-positive cocci or bacilli, catalase negative and not endospore producing.
Select two colonies that, due to the characteristics analyzed, may belong to different types
of lactic bacteria.
1.5 Isolate each of the two selected strains in pure culture by successive passages
(at least 3) by non-selective media (MRS without antibiotic). Between each passage,
incubate the plates at isolation temperature.
1.6. After isolation, confirm cell morphology, gram character, absence of catalase and
endospores. Refer to the strains with the code LACxA and LACxB where x is the student
group number.
2. Identification
2.1. For the strain of your group that is selected in the class, carry out its identification at
species level, using the miniaturized galleries API 50CH and the suspension medium
API 50CHL, according to the manufacturer's instructions.
2.3 If it is impossible to to achieve an identification, check that the final isolation plate does
not contain a mixed culture (failure of the purification process).
REF 50 300 07945F - GB - 2002/11
®
50 CH IVD
Carbohydrates
bioMérieux® sa English - 1
api® 50 CH 07945F - GB - 2002/11
REAGENTS AND MATERIAL REQUIRED BUT NOT SPECIMENS (COLLECTION AND PREPARATION)
PROVIDED API 50 CH is not for use directly with clinical or other
Reagents : specimens.
- Inoculation medium : The microorganisms to be identified must first be isolated
API 50 CHL Medium (Ref. 50 410) on a suitable culture medium according to standard
API 50 CHB/E Medium (Ref. 50 430) microbiological techniques.
(+ products mentioned in the package inserts
of these media) INSTRUCTIONS FOR USE
or any other suitable medium Depending on which medium is used, API 50 CHL
- Mineral oil (Ref. 70 100) Medium or API 50 CHB/E Medium, carefully read the
- McFarland Standard (Ref. 70 900) or corresponding package insert.
DENSIMAT (Ref. 99 234) or ATB® Densitometer
- Identification software (consult bioMérieux) Preparation of the strips
Each strip is made up of 5 smaller strips each containing
Material :
10 numbered tubes.
- Pipettes or PSIpettes
• Prepare an incubation box (tray and lid).
- Ampule rack
• Record the reference of the strain on the elongated flap
- Ampule protector
of the tray. (Do not record the reference on the lid as it
- Swabs
may be misplaced during the procedure.)
- General microbiology laboratory equipment
• Distribute about 10 ml of distilled water or de-
mineralized water [or any water without additives or
WARNINGS AND PRECAUTIONS chemicals which may release gases (e.g. Cl2, CO2, etc.)]
• For in vitro diagnostic use and microbiological into the honeycombed wells of the tray to create a
control. humid atmosphere.
• For professional use only. • Remove the 2 strips (0-19 and 20-39) from their
• This kit contains products of animal origin. Certified packaging, separate into 4 smaller strips (0-9, 10-19,
knowledge of the origin and/or sanitary state of the 20-29 and 30-39) and place all 4 in the incubation tray.
animals does not totally guarantee the absence of • Take the remaining smaller strip (40-49) out of the
transmissible pathogenic agents. It is therefore packaging and place it next to the others in the
recommended that these products be treated as incubation tray to complete the strip.
potentially infectious, and handled observing the usual
safety precautions (do not ingest or inhale). Preparation of the inoculum
• All specimens, microbial cultures and inoculated • Culture the microorganism on a medium adapted to its
products should be considered infectious and handled growth.
appropriately. Aseptic technique and usual precautions • Check the purity of the strain.
for handling the bacterial group studied should be • Harvest all the bacteria from a solid medium using a
observed throughout this procedure. Refer to "NCCLS swab or from a liquid medium using centrifugation.
M29-A, Protection of Laboratory Workers from • Prepare the inoculum in the appropriate medium (see
Instrument Biohazards and Infectious Disease the API 50 CHL Medium and API 50 CHB/E Medium
Transmitted by Blood, Body Fluids, and Tissue; package inserts).
Approved Guideline - December 1997". For additional This suspension must be used immediately after
handling precautions, refer to "Biosafety in preparation.
Microbiological and Biomedical Laboratories, HHS
Inoculation of the strips
Publication No. (CDC) 93-8395, 3rd Edition (May
1993)", or to the regulations currently in use in each Distribute the bacterial suspension using a sterile pipette
country. into the 50 tubes as follows :
• Do not use reagents past the expiration date. • Tilt the incubation box slightly forwards.
• Before use, check that the packaging of the various • Avoid the formation of bubbles by placing the tip of the
components is intact. pipette against the side of the cupule.
• Do not use strips which have been damaged : cupules • When only the tube is to be inoculated, do not exceed
deformed, desiccant sachet open, ... the top of the tube so as to maintain anaerobic
• The performance data were obtained using the conditions.
procedure indicated in this package insert. Any change • When the tube and the cupule are to be completely
or modification in the procedure may affect the results. filled, avoid the formation of a concave or convex
• Interpretation of the test results should be made taking meniscus.
into consideration the patient history, the source of the • Incubate the strips at the optimum temperature for
specimen, colonial and microscopic morphology of the growth of the group of microorganisms being tested :
strain and, if necessary, the results of any other tests 30°C, 37°C or 55°C.
performed, particularly the antimicrobial susceptibility
patterns.
STORAGE CONDITIONS
The strips should be stored at 2-8°C until the expiration
date indicated on the packaging.
bioMérieux® sa English - 2
api® 50 CH 07945F - GB - 2002/11
READING AND INTERPRETATION The results form a biochemical profile which, when
Reading the strips entered in the identification software, provides the
identification of Lactobacillus and related genera or
The strips are read after the stipulated incubation times
Bacillus and related genera, Enterobacteriaceae and
(e.g., 24 hrs., 48 hrs.), depending on the microorganism
Vibrionaceae.
and the type of reaction studied.
NOTE : The results may be used for other purposes :
Interpretation • Epidemiological grouping into types of the
Interpret each test (positive (+), negative (-), doubtful (?)) microorganism.
and record the results on the result sheet. • Taxonomical analysis of a group of microorganisms.
• Classification of an unknown bacterial population into
homogeneous groups.
QUALITY CONTROL
The strips are systematically controlled at various stages of their manufacture. For those users who wish to perform their
own quality control tests with the strip, the following strains may be used :
For Lactobacillus : Lactobacillus paracasei ssp paracasei NFCB 206 or ATCC BAA-52 (with API 50 CHL Medium)
0 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49
24 - - - - - + - - - - + + + + - - V - + V - + + - V + V V + - - + + + + - - - - - + - + - - - - V - -
48 - - - - - + - - - - + + + + - - + - + + - + + V + + + + + - - + + + + - - - - V + - + - - - - + - -
For Bacillus : Bacillus polymyxa (*) ATCC 43865 (with API 50 CHB/E Medium)
0 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49
24 - + - - + + + - - V + + + + - V - - + - - + - + + + + + + + + + + - + + + + - + + - - - - - - - - -
48 - + - - + + + - - + + + + + - V - - + - - + - + + + + + + + + + + V + + + + - + + - - - - - - - - -
(*) Bacillus polymyxa identified as Paenibacillus polymyxa with API 50 CH and API 50 CHB/E Medium.
Results obtained after incubation at 30°C.
For Enterobacteriaceae : Klebsiella pneumoniae ssp pneumoniae ATCC 35657 (with API 50 CHB/E Medium)
0 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49
24 - + - - + + + - + - + + + + - + - + + + - + + - + + + + + + + + + - - + + - - + - - - - + + - + + -
48 - + - V + + + - + - + + + + - + - + + + - + + V + + + + + + + + + - - + + - - + V - - - + + - + + -
ATCC : American Type Culture Collection, 10801 University Boulevard, Manassas, VA 20110-2209, USA.
NCFB : National Collection for Food Bacteria (=NCDO), Institute of Food Research, Reading Laboratory, Earley Gate, Reading
RG6 6BZ, ENGLAND
It is the responsibility of the user to perform Quality Control in accordance with any local applicable regulations.
LITERATURE REFERENCES p. I
INDEX OF SYMBOLS p. II
®
50 CH IVD
Carbohidratos
bioMérieux® sa Português - 1
api® 50 CH 07945F - PT - 2002/11
CONDIÇÕES DE ARMAZENAMENTO
As galerias conservam-se a 2º-8ºC até à data de validade
indicada na embalagem.
bioMérieux® sa Português - 2
api® 50 CH 07945F - PT - 2002/11
Para Bacillus : com a estirpe/cepa Bacillus polymyxa (*) ATCC 43865 (com API 50 CHB/E Medium)
0 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49
24 - + - - + + + - - V + + + + - V - - + - - + - + + + + + + + + + + - ++ + + - + + - - - - - - - - -
48 - + - - + + + - - + + + + + - V - - + - - + - + + + + + + + + + + V + + + + - + + - - - - - - - - -
(*) Bacillus polymyxa identificado a Paenibacillus polymyxa em API 50 CH e API 50 CHB/E Medium.
Resultados obtidos após incubação a 30°C.
Para Enterobacteriaceae : com a estirpe/cepa Klebsiella pneumoniae ssp pneumoniae ATCC 35657 (com API 50
CHB/E Medium)
0 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49
24 - + - - + + + - + - + + + + - + - + + + - + + - + + + + + + + + + - - + + - - + - - - - + + - + + -
48 - + - V + + + - + - + + + + - + - + + + - + + V + + + + + + + + + - - + + - - + V - - - + + - + + -
ATCC : American Type Culture Collection, 10801 University Boulevard, Manassas, VA 20110-2209, USA.
NCFB : National Collection for Food Bacteria (=NCDO), Institute of Food Research, Reading Laboratory, Earley Gate, Reading
RG6 6BZ, ENGLAND
É da responsabilidade do utilizador assegurar que o controlo de qualidade é efectuado em conformidade com a
legislação local em vigor.
LIMITES DO TESTE COMPORTAMENTO FUNCIONAL
• Qualquer identificação de espécies não enumeradas Consultar o comportamento funcional dos meios
nas bases de dados API 50 CHL e API 50 CHB/E está associados a esta galeria.
sob a responsabilidade do utilizador.
ELIMINAÇÃO DE RESÍDUOS
• Devem apenas ser utilizadas culturas puras contendo
um único tipo de microrganismo. É da responsabilidade de cada laboratório gerir os
resíduos e os efluentes que este produz consoante a sua
RESULTADOS ESPERADOS natureza e o seu perigo, e assegurar (ou fazer assegurar)
Consultar o Quadro de Identificação no final deste folheto o tratamento e a eliminação em conformidade com as
informativo para saber os resultados esperados para as regulamentações aplicáveis.
diferentes reacções bioquímicas.
BIBLIOGRAFIA p. I
QUADRO DE SÍMBOLOS p. II
Brasil: Distribuído por biolab-Mérieux, S.A. - Estrada do Mapuá, 491 - Jacarepaguá - R.J. - CEP 22710-261
CNPJ: 33.040.635/0001-71
Atendimento ao Consumidor Tel.: 0800-264848
Prazo de Validade, N° de Lote, N° de Registro de Ministério da Saúde e Responsável Técnico:
VIDE EMBALAGEM
®
50 CHL Medium IVD
bioMérieux SA English - 1
api® 50 CHL Medium 07486H - en - 2011/07
QUALITY CONTROL
The media and strips are systematically controlled at various stages of their manufacture. For those users who wish to
perform their own quality control tests with the strip, it is preferable to use the strain 1. Lactobacillus plantarum
®
ATCC 14917 or the following strain :
2. Lactobacillus paracasei ssp paracasei NCFB 206 or ATCC BAA-52
ATCC : American Type Culture Collection, 10801 University Boulevard, Manassas, VA 20110-2209, USA.
NCFB : National Collection for Food Bacteria (=NCDO), Institute of Food Research, Reading Laboratory, Earley Gate, Reading
RG6 6BZ, ENGLAND
0 0 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49
1. 24 – – – – + + – – – – + + + + – – – – + – – – + + + + + + + – + + + – + – – – – + + – – – – – – + – –
48 – – – – + + – – – – + + + + – – – – + + + – + + + + + + + V + + + – + + – – – + + – – – – – – + – –
2. 24 – – – – – + – – – – + + + + – – V – + V – + + – V + V V + – – + + + + – – – – – + – + – – – – V – –
48 – – – – – + – – – – + + + + – – + – + + – + + V + + + + + – – + + + + – – – – V + – + – – – – + – –
It is the responsibility of the user to perform Quality Control in accordance with any local applicable regulations.
bioMérieux SA English - 2
api® 50 CHL Medium 07486H - en - 2011/07
WARRANTY
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including any implied warranties of MERCHANTABILITY
AND FITNESS FOR A PARTICULAR USE. bioMérieux
shall not be liable for any incidental or consequential
damages. IN NO EVENT SHALL BIOMERIEUX’S
LIABLITY TO CUSTOMER UNDER ANY CLAIM
EXCEED A REFUND OF THE AMOUNT PAID TO
BIOMERIEUX FOR THE PRODUCT OR SERVICE
WHICH IS THE SUBJECT OF THE CLAIM.
PROCEDURE p. I
IDENTIFICATION TABLE p. II
LITERATURE REFERENCES p. III
INDEX OF SYMBOLS p. IV
BIOMERIEUX, the blue logo, API, ATB and apiweb are used, pending and/or registered trademarks belonging to bioMérieux SA or one of its subsidiaries.
CLSI is a trademark belonging to Clinical and Laboratory Standards Institute, Inc.
ATCC is a trademark belonging to American Type Culture Collection.
Any other name or trademark is the property of its respective owner.
Lactobacillus
samtliga bakterier
alle bakterier
API Suspension Medium
wszystkie bakterie
2 ml
2n 2 McF
API
Suspension
Medium
5 ml
2 McF
API 50 CH
+ - + - + -
bioMérieux SA I
api® 50 CHL Medium 07486H - xl - 2011/07
TABLEAU D'IDENTIFICATION / IDENTIFICATION TABLE / PROZENTTABELLE / TABLA DE IDENTIFICACION / TABELLA DI IDENTIFICAZIONE / QUADRO DE IDENTIFICAÇÃO
/ IDENTIFIERINGSTABELL / IDENTIFIKATIONSTABEL / TABELA IDENTYFIKACYJNA
% de réactions positives après 48 H (± 6 h) à 36°C ± 2°C / % of positive reactions after 48 hrs. (± 6 hrs) at 36°C ± 2°C / % der positiven Reaktionen nach 48 Std. (± 6 Std) bei 36°C ± 2°C /% de las reacciones positivas después de 48H
(± 6 H) a 36°C ± 2°C / % di reazioni positive dopo 48 ore (± 6 ore) a 36°C ± 2°C / % das reacções positivas após 48 h (± 6 h) a 36°C ± 2°C / % 48 (± 6 ) 36°C ± 2°C /
% positiva reaktioner efter 48 tim. (± 6 tim) vid 36°C ± 2°C / % positive reaktioner efter 48 timer (± 6 timer) ved 36°C ± 2°C / % pozytywnych reakcji po 48 godzinach (± 6 godz.) w 36°C ± 2°C
bioMérieux SA II
api® 50 CHL Medium 07486H - xl - 2011/07
1. BAYER A.S., CHOW A.W., BETTS D., GUZE L.B. 8. SHARPE M.E., HILL L.R., LAPAGE S.P.
Lactobacillemia-Report of Nine Cases. Pathogenic Lactobacilli.
Important Clinical and Therapeutic Considerations. (1973) J. Med. Microbiol. 6, 281-286.
(1978) Am. J. of Med. 64, 808-813. 9. SNEATH P.H.A., MAIR N.S., SHARPE E., HOLT J.G.
2. DE MAN JC., ROGOSA M., SHARPE ME. Bergey's Manual of Systematic Bacteriology
A medium for the Cultivation of Lactobacilli. (1986) Williams and Wilkins - Vol 2.
(1960) J. Appl. Bact. 23, 130-135. 10. FLEET G.H., LAFON-LAFOURCADE S., RIBEREAU-
3. LATORRE-GUZMAN B.A., KADO C.I., KUNKEE R.E. GAYON P.
Lactobacillus hordniae, a New Species from the Leafhopper Evolution of Yeasts and Lactic Acid Bacteria During
(Hordnia circellata). Fermentation and Storage of Bordeaux Wines.
(1977) Int. J. Syst. Bact. 27, 362-370. (1984) Appl. Environ. Microbiol. 48, 1034-1038.
4. LAUDAT P., PENEAU M., PINON G., LANSON Y., 11. HOFER F.
AUDURIER A. Identifizierung von Milchsäurebakterien mit Hilfe des API-
Pyélonéphrite et Septicémie à Lactobacillus acidophilus. Systems.
(1982) Méd. et Mal. Infect. 12, 289-291. (1976) Schweiz. Milchw. Forsch. 5, 17-22.
5. LE MINOR L., VERON M. 12. LABAN P., FAVRE C., RAMET F., LARPENT J.P.
Bactériologie Médicale. Lactobacilli isolated from French saucisson (Taxonomic
2° Edition. Study).
(1989) Flammarion Médecine Sciences. (1978) Zbl. Bakt. Hyg.I. Aby. Orig. B, 166, 105-111.
6. MURRAY P.R., BARON E.J., JORGENSEN J.H., 13. MARET R., SOZZI T.
PFALLER M.A., YOLKEN R.H. Flore lactique de fromageries d'alpages suisses
Manual of Clinical Microbiology. (1976) Le Lait, 56, 1-13.
th
8 Edition.
(2003) American Society for Microbiology, Washington, D.C.
7. ROGOSA M., SHARPE M.E.
An approach to the classification of the Lactobacilli.
(1959) J. Appl. Bact. 22, 329-340.
bioMérieux SA III
api® 50 CHL Medium 07486H - xl - 2011/07
Se handhavandebeskrivningen / Se brugsanvisning
Sprawd w instrukcji obs ugi
Contenu suffisant pour "n" tests
Contains sufficient for <n> tests
Inhalt ausreichend für <n> Prüfungen
Contenido suficiente para <n> ensayos
Contenuto sufficiente per "n" saggi
Conteúdo suficiente para “n” ensaios
« »
Räcker till "n" antal tester / Indeholder tilstrækkeligt til "n" test
Wystarczy na wykonanie <n> testów
®
50 CHL Medium IVD
Lactobacillus e semelhantes
bioMérieux SA Português - 1
api® 50 CHL Medium 07486H - pt - 2011/07
CONTROLO DE QUALIDADE
Os meios e galerias são sujeitos a controlos de qualidade sistemáticos nas diferentes etapas do seu fabrico. Além disso,
o utilizador pode efectuar um controlo bacteriológico dos testes da galeria, preferencialmente com a estirpe/cepa
®
1. Lactobacillus plantarum ATCC 14917 ou com a estirpe/cepa seguinte :
2. Lactobacillus paracasei ssp paracasei NCFB 206 ou ATCC BAA-52
ATCC : American Type Culture Collection, 10801 University Boulevard, Manassas, VA 20110-2209, USA.
NCFB : National Collection for Food Bacteria (=NCDO), Institute of Food Research, Reading Laboratory, Earley Gate, Reading
RG6 6BZ, ENGLAND
0 0 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49
1. 24 – – – – + + – – – – + + + + – – – – + – – – + + + + + + + – + + + – + – – – – + + – – – – – – + – –
48 – – – – + + – – – – + + + + – – – – + + + – + + + + + + + V + + + – + + – – – + + – – – – – – + – –
2. 24 – – – – – + – – – – + + + + – – V – + V – + + – V + V V + – – + + + + – – – – – + – + – – – – V – –
48 – – – – – + – – – – + + + + – – + – + + – + + V + + + + + – – + + + + – – – – V + – + – – – – + – –
bioMérieux SA Português - 2
api® 50 CHL Medium 07486H - pt - 2011/07
PROCEDIMENTO p. I
QUADRO DE IDENTIFICAÇÃO p. II
BIBLIOGRAFIA p. III
QUADRO DE SÍMBOLOS p. IV
A BIOMERIEUX, o logotipo azul, API, ATB e apiweb são marcas utilizadas, depositadas e/ou registadas, propriedade exclusiva da bioMérieux SA ou de
uma das suas filiais.
CLSI é uma marca propriedade exclusiva da Clinical and Laboratory Standards Institute, Inc.
ATCC é uma marca propriedade exclusiva da American Type Culture Collection.
As outras marcas e nomes de produtos mencionados neste documento são marcas comerciais dos seus proprietários respectivos.
Brasil: Distribuído por bioMérieux Brasil, S.A. - Estrada do Mapuá, 491 - Jacarepaguá - R.J. - CEP 22710-261
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Prazo de Validade, N° de Lote, N° de Registro de Ministério da Saúde e Responsável Técnico:
VIDE EMBALAGEM
Lactobacillus
samtliga bakterier
alle bakterier
API Suspension Medium
wszystkie bakterie
2 ml
2n 2 McF
API
Suspension
Medium
5 ml
2 McF
API 50 CH
+ - + - + -
bioMérieux SA I
api® 50 CHL Medium 07486H - xl - 2011/07
TABLEAU D'IDENTIFICATION / IDENTIFICATION TABLE / PROZENTTABELLE / TABLA DE IDENTIFICACION / TABELLA DI IDENTIFICAZIONE / QUADRO DE IDENTIFICAÇÃO
/ IDENTIFIERINGSTABELL / IDENTIFIKATIONSTABEL / TABELA IDENTYFIKACYJNA
% de réactions positives après 48 H (± 6 h) à 36°C ± 2°C / % of positive reactions after 48 hrs. (± 6 hrs) at 36°C ± 2°C / % der positiven Reaktionen nach 48 Std. (± 6 Std) bei 36°C ± 2°C /% de las reacciones positivas después de 48H
(± 6 H) a 36°C ± 2°C / % di reazioni positive dopo 48 ore (± 6 ore) a 36°C ± 2°C / % das reacções positivas após 48 h (± 6 h) a 36°C ± 2°C / % 48 (± 6 ) 36°C ± 2°C /
% positiva reaktioner efter 48 tim. (± 6 tim) vid 36°C ± 2°C / % positive reaktioner efter 48 timer (± 6 timer) ved 36°C ± 2°C / % pozytywnych reakcji po 48 godzinach (± 6 godz.) w 36°C ± 2°C
bioMérieux SA II
api® 50 CHL Medium 07486H - xl - 2011/07
1. BAYER A.S., CHOW A.W., BETTS D., GUZE L.B. 8. SHARPE M.E., HILL L.R., LAPAGE S.P.
Lactobacillemia-Report of Nine Cases. Pathogenic Lactobacilli.
Important Clinical and Therapeutic Considerations. (1973) J. Med. Microbiol. 6, 281-286.
(1978) Am. J. of Med. 64, 808-813. 9. SNEATH P.H.A., MAIR N.S., SHARPE E., HOLT J.G.
2. DE MAN JC., ROGOSA M., SHARPE ME. Bergey's Manual of Systematic Bacteriology
A medium for the Cultivation of Lactobacilli. (1986) Williams and Wilkins - Vol 2.
(1960) J. Appl. Bact. 23, 130-135. 10. FLEET G.H., LAFON-LAFOURCADE S., RIBEREAU-
3. LATORRE-GUZMAN B.A., KADO C.I., KUNKEE R.E. GAYON P.
Lactobacillus hordniae, a New Species from the Leafhopper Evolution of Yeasts and Lactic Acid Bacteria During
(Hordnia circellata). Fermentation and Storage of Bordeaux Wines.
(1977) Int. J. Syst. Bact. 27, 362-370. (1984) Appl. Environ. Microbiol. 48, 1034-1038.
4. LAUDAT P., PENEAU M., PINON G., LANSON Y., 11. HOFER F.
AUDURIER A. Identifizierung von Milchsäurebakterien mit Hilfe des API-
Pyélonéphrite et Septicémie à Lactobacillus acidophilus. Systems.
(1982) Méd. et Mal. Infect. 12, 289-291. (1976) Schweiz. Milchw. Forsch. 5, 17-22.
5. LE MINOR L., VERON M. 12. LABAN P., FAVRE C., RAMET F., LARPENT J.P.
Bactériologie Médicale. Lactobacilli isolated from French saucisson (Taxonomic
2° Edition. Study).
(1989) Flammarion Médecine Sciences. (1978) Zbl. Bakt. Hyg.I. Aby. Orig. B, 166, 105-111.
6. MURRAY P.R., BARON E.J., JORGENSEN J.H., 13. MARET R., SOZZI T.
PFALLER M.A., YOLKEN R.H. Flore lactique de fromageries d'alpages suisses
Manual of Clinical Microbiology. (1976) Le Lait, 56, 1-13.
th
8 Edition.
(2003) American Society for Microbiology, Washington, D.C.
7. ROGOSA M., SHARPE M.E.
An approach to the classification of the Lactobacilli.
(1959) J. Appl. Bact. 22, 329-340.
bioMérieux SA III
api® 50 CHL Medium 07486H - xl - 2011/07
Se handhavandebeskrivningen / Se brugsanvisning
Sprawd w instrukcji obs ugi
Contenu suffisant pour "n" tests
Contains sufficient for <n> tests
Inhalt ausreichend für <n> Prüfungen
Contenido suficiente para <n> ensayos
Contenuto sufficiente per "n" saggi
Conteúdo suficiente para “n” ensaios
« »
Räcker till "n" antal tester / Indeholder tilstrækkeligt til "n" test
Wystarczy na wykonanie <n> testów
ANNEX
Extracts from
Tenreiro, R. (1995). Polyphasic taxonomic analysis in Leuconostoc oenos. PhD thesis.
Universidade de Lisboa. [In Portuguese]
including:
1.2 - Isolation of wine lactic acid bacteria: fundamentals and processes
1.3 - Isolation method
1.4 - Strain selection profiles, presumptive identification and representativeness
1.5 - Storage processes
1.6 - Growth conditions, culture media and optimisation
including:
2.1 - One genus, but which?
2.2 - Isolating Leuconostoc spp.