STAPHYLOCOCCUS

AUREUS
Dr Surender Kaur
Asst.Prof. GMC
 Family: Micrococcaceae
 Genera:
1) Staphylococcus
2) Micrococcus
3) Planococcus
4) Stomatococcus
 Difference between:
character Staphylo Micro Plano
Arrangement clusters Irreg/cluster/tetrad tetrad
Anaerobic + - -
Teichoic acid + - -
pigmentation + - -
Glucose F O
fermentation
lysostaphin S R R
bacitracin R S
catalase + +
oxidase - +
 Characterstics of staphylococcus
 Aerobic/facultative anaerobe
 Arrangement: pairs, tetrads, cluster
 Catalase +ve
 Oxidase –ve
 Coagulase +ve
 On basis of coagulase Test:
Coagulase Positive Coagulase Negative
Staphylococcus aureus S.epidermidis S.schleiferi

S.haemolyticus S.lugdunesis

S.saprophyticus

S.sacrolyticus

S.waeneri

S.hominis

S.capitus
 Morphology:
 GPC
 Nonmotile, nonsporing, noncapsulated
 Grape like cluster
 Culture:
 Aerobic /facultative anaerobe
 Temp- 10-42 deg C
 PH- 7.4-7.6
Nutrient agar- Blood agar-
Round, Convex, β- haemolysis
2-4mm, butyrous, glistening
surface, golden yellow
nondiffusible pigment

Nutrient agar slope- Selective Media-

Oil paint Mannitol salt agar
Salt Milk agar
B.A- S.aureus MSA- S.aureus
 Virulence Factor –
 Cell wall associated factors
 Enzymes
 Toxin
I. Cell wall associated factor-
1. Peptidoglycan
2. Teichoic acid
3. Protein A
1. Peptidoglycan-
 Confers rigidity and integrity to cell wall.
 Activates the complement pathway and releases
inflammatory cytokines.
2. Teichoic acid-
 Antigenic in nature and facilitates adhesion on host
surface.
 Protects bacteria from complement mediated opsonisation.
3. Protein A-
 Present in S.aureus have chemotactic, anti-inflamatory,
anti-phagocytic property.
 It binds with Fc portion of IgG leaving Fab free to combine
with specific antigen.
 Coagglutination-
Use- for Streptococcal grouping and Gonococcal typing.
III. Enzymes-
1. Coagulase –
 Two type- bound coagulase and Coagulase enzyme.
 Bound coagulase (clumping factor) is surface protein
responsible for Slide coagulase Test.
 Coagulase enzyme brings about clotting of human
plasma or rabbit plasma. It acts with coagulase reacting
factor (CRF) in plasma, converts fibrinogen in to fibrin .
It occurs in Tube coagulase.
II Other enzymes-
Nuclease- heat stable DNAse characterstic of S.aureus
Hyluraunidase- helps in spreading of infection
DN Ase Test
III. Toxins-
i. Cytolytic toxin- haemolysins
ii. Panton Valentine Leucocidin (PVL)
iii. Enterotoxin
iv. Toxic shock syndrome toxin
v. Exfoliative toxin
 Enterotoxin-
 Preformed toxins responsible for staph. Food poisoning.
 Symptoms- N,V and D
 Inc. period- after 2-6 hrs of food cosumption.
 Predominently grows on – meat, fish, milk and product.
 Serotyping shows- Eight types of enterotoxins.
 Site of action- on Autonomic nervous system.
 Detection of toxin- ELISA, Latex agglutination.
 Toxic sock syndrome toxin (TSST)-
 Fatal multisystem disease.
 Associated with infection of mucosa by TSST-type I
produced by S.aureus
 Present as fever, myalgia, N ,V, Rash Hypotension.
 Pathogenesis- TSST-I absorbed in to circulation from
abraded mucosa during menstruation and superantigen
acts by nonspecific stimulation of T- lymphocytes ( Vβ
region of T cell receptor ) causing excess production of
cytokines which leads to fatal multiorgan involment.
 SuperAntigens-
 Ag that activate a large fraction of the T cells, setting off
massive immune response. These antigens are called Super
Ag.
 EX. Staphylococcal enterotoxins and TSST
 A typical antigen must be processed by an APC, after which it binds
to both the α and β chain of the TCR.
 Superantigens don’t require processing and do not bind to the α
chain. Instead, they link the β chain of the TCR directly to the
class II MHC molecule on the APC.

 Exfoliative Toxin-
 It is responsible for Staphylococcal scalded skin syndrome.
 Occurs- in Newborn and adults.
 Symptoms- blisters and bullae formation which exfolites and
spreads.
 Severe form in newborn is Ritters disease
 Milder form- pemphigous or impetigo.
 PATHOGENESIS

Colonization-
most common site of colonization is anterior nares, axilla, perineal
skin and serve as reservoir for future infection.
Introduction in tissue-
 through abraded skin or instrumentation.
Invasion-
through enzymes
Clinical manifestation-

Skin & Musculos Resp Bacterem UTI Toxin

soft keletal tract ia mediated
tissue
Folliculiti Septic VAP Sepsis TSS
s arthritis
Furuncle osteomye Septic shock Food pg
litis pulmonar
y emboli
Carbuncle Abscess Pneumoth Infective
orax endocardi
tis
Impetigo Empyema
Cellulitis
 Lab Diagnosis-
 Sample collection
 Direct Microscopy
 Culture-
 Culture smear microscopy- GPC (1μ ) in clusters.
 Biochemical Tests for Identification-
1. Catalase Test-
2. H-F oxi-ferm Test- Staph- Fermentative
Micro-Oxidative
3. Coagulase Test- To differentiate S.aureus from CONS
4. DNAse Test- Confirmation test for S.aureus
5. Phosphatase Test- Positive for S.aureus and
S.epidermidis.
 Typing of S.aureus-
Phenotyping- Bacteriophage typing and Antibiogram typing
Genotyping- PCR, Ribotyping
 Bacteriophage typing-

 National Referrence center for S.aureus phage typing-

MAMC NewDehli.
Drug Resistance-
1. Production of β-Lactamase enzyme:
 Plasmid mediated resistance.
 Transferred between strains of S.aureus by Transduction.
 Resistance can be overcomed by- β-Lactamase inhibitors
like clavulanic acid
2. By alteration of Penicillin binding protein- MRSA
MRSA- It is mediated by chromosomally coded gene called
mec A gene which alters PBP to PBP-2a.
Mech. Action-PBP is essential protein for cell wall synthesis.
Β-Lactam drug binds with this protein and inhibits cell wall
synthesis.
BORSA- Boderline oxacillin resistant S.aureus
 Low level resistance to oxacillin which is mediated by non-
mec A gene.
 It is due to hyperproduction of Β-Lactamase enzyme.
 TYPES OF MRSA
CA- MRSA HA- MRSA
Community associated MRSA Hospital associated MRSA
These strain express mecA gene These strain express mecA gene
subtype IV, V, VI subtype I, II, III
MORE VIRULENT less
Produces toxin like panton They are MDR
vallentine toxin
Causes- invasive skin and soft Responsible for nosocomial
tissue infection outbreaks
 DETECTION OF MRSA
1. Disc Diffusion test- by using Cefoxitin or Oxacillin disc.
2. Oxacillin screening agar- (6μg) and 2-4% NaCl containing
media.
3. PCR- mecA gene detection.
 Drug of choice in MRSA- Vancomycin
VISA and VRSA-
 Rare in India
 Mechanism- by Van A and B gene
Control Measures-
1. Hand washing
2. Screening for MRSA carriers
3. Treatment of carriers- Nasal carrier- topical mupirocin and
skin carrier-chlorhexdine
4. Fumigation
 COAGULASE NEGATIVE S.AUREUS (CONS)
Commensal flora on skin, oropharynx and vagina.
 S.epidermidis, saprophyticus, lugdenensis and sheilferi
Pathogenesis of S.epidermidis-
1. Intial adhesion on prosthetic devices- through fibrinogen
2. Colonization- It produces glycocalyx or slime which forms biofilm
on surface of device. Biofilm act as protective barrier of
bacteria from host defence mechanism.
3. Manifestation- s.epidemidis causes prosthetic device related
diseases ex- valvular endocardittis , shunt infection, stich
abseccess.
S.saprophyticus-
 Causes UTI due to haemagglutinin or adhesin protein.
 Differentiated from S.aureus by RESISTANT to NOVOBIOCIN
Thankyou