TEST FOR IDENTIFYING

THE ALPHA-HEMOLYTIC
STREPTOCOCCI
OPTOCHIN TEST/ TAXO P DISK
Differentiate Streptococcus pnemoniae from other
Alpha hemolytic Strep like Stretococcus mitis.

In the presence of Optochin, the colonies of S.

pnumoniae are selectively lysed
OPTOCHIN TEST/ TAXO P DISK

Optochin other name ethylhydrocupreine

hydrochloride

The optochin test the fragility of the bacterium cell

membrane,the optochin can inhibit the S. pnuemonaie
while S. mitis cannot.
BILE SOLUBILITY TEST
• Use of bile salt (Sodium deoxycholate) can cause lysis
to some bacteria like Strep. Pnuemoniae

• Positive: S. pnuemoniae
• Negative: Enterobacter faecalis
TEST FOR IDENTIFICATION
OF BETA-HEMOLYTIC
STREPTOCOCCI
Bacitracin-SXT susceptibility
It is for the identification of group A strep and Group B beta
hemolytic streptococci

Group A streptococci are susceptible to bacitracin but resistant to

SXT.
Group B beta hemolytic strep are resistant to both bacitracin and SXT

Any zone of inhibition around either disk indicates that the organism
is susceptible
• For bacitracin test

• POSITIVE(+) s. pyogenes
• NEGATIVE(-) S. agalactiae
PYRase test(PYR test)
• Differentiate enterococcus species from non-
enterococcus species
• Also Group A strep are positive in the pyrase test
• PYRase test hydrolyzed PYR producing a red color
upon addition of p-dimethylcinnamaldehyde
PYRase test(PYR test)

• This test is used for the detection of pyrolidonyl arylamidase also

called as pyrolidonyl aminopeptidase(PYR) which produce by some
Enterobacteriaciae and Streptococcus pyogenes
CAMP
• Identify group B streptococcus

• Group B strep produces extracellular protein that

enhance hemolysis of beta hemolytic S. aureus on SBA

• Strep agalactiae have a charateristics of arrowhead

hemolytic pattern
CAMP test
• POSITIVE: produces a arrowhead pattern hemolysis

ex. Strep agalactiae

NEGATIVE: no zone of enhanced hemolysis
ex. pyogenes
Hippurate hydrolysis
• Identify group B streptococcus alternative to CAMP
• Organisms that produce hippuricase(Hippurate
hydrolase) hydrolyze sodium Hippurate to benzoate
and glycine.
Hippurate hydrolysis
• POSITIVE: purple color after adding ninhydrin reagent
ex. Strep agalactiae
• NEGATIVE: no color change
ex. Strep pyogenes
Slide agglutination tests
• Latex particles coated with group specific antibodies agglutinate in
presence of bacterial antigens
• Organisms agglutinate in corresponding anti-sera
Hemolytic Reaction Lancefield Group Species
Alpha none S.pnuemoniae
Beta A s.Pyogenes
B s. Agalactiae
C S. Dysagalactiae
Gamma D S.Bovis
Alpha,beta, or gamma D Enterococcus spp
Alpha and gamma None Viridans strep
Small colony variants of A,C,F,G or Anginous group
ungroupable
TESTS FOR IDENTIFICATION OF NON-
HEMOLYTIC
STREPTOCOCCI/ENTEROCOCCI
PYRASE TEST
BILE ESCULIN AGAR
• Distinguish Group D Streptococcus and Enterococcus
from other lancefield group

• Group D streptococcus can grow in 40% bile and

hydrolize esculin to formed esculetin and this esculetin
reacts to ferric citrate forming a brown-black
precipitate
• POSITIVE: Growth with Blackening
6.5% Salt broth/ Salt tolerance test
• Differentiate Group D streptococci from
nonenterococci
• Group D enterococci are salt tolerant Group D
nonenterococci are susceptible
 BILE ESCULIN 6.5% salt Pyrase
broth
Group D pos pos pos
enterococcus
Group D non pos nega nega
enterococcus
Not Group D nega nega nega
TEST FOR IDENTIFYING NEISSERIA
SPECIES
CYSTINE TRYPICASE AGAR
-Differentiate Neisseria species by using different carbohydrate suspend
in the CTA tube.
- The bacteria oxidizes the different types of carbohydrate changing the
color of phenol red into yellow color.
 Glucose Maltose Lactose Sucrose
N. pos neg neg neg
gonorrhoeae
N. pos pos neg neg
Meningitidis
N. Lactamica pos pos pos neg
N. Sicca pos pos neg pos
N. Elongata neg neg neg neg