Catalase-Negative, Gram- Differentiation is based on:

Positive Cocci:  Cell wall structure

Streptococcus,  Hemolytic patterns of BAP
Enterococcus, and Similar  Reaction of antibodies to
specific
bacterial antigens
 Lancefield Classification
scheme
 Biochemical identification

Most commonly encountered

in human
infections include:
▪ Streptococcus pyogenes
▪ Streptococcus agalactiae
▪ Streptococcus pneumoniae
▪ Enterococcus faecalis
▪ Enterococcus faecium
▪ Viridans streptococci group

Organisms General Characteristics

Introduction  Streptococcus and
- The enterococci, formerly Enterococcus spp. belong to
known as group D streptococci, the family Streptococcaceae
have been classified in their  Members of both genera are
own genus, Enterococcus catalasenegative, gram-
- Similarly, lactococci, positive cocci that are
previously classified as group N usually arranged in pairs or
streptococci, now belong in the chains.
genus Lactococcus  A negative catalase test
- Although traditional result
phenotypic differentiates streptococci and
characteristics such as enterococci from staphylococci.
hemolysis and Lancefield  Weak false-positive catalase
classification (antigen reactions can be seen when
serogrouping) are still useful in growth is taken from media
presumptive ID, nucleic acid containing blood, owing to
studies have provided more the peroxidase activity of
information on the genetic hemoglobin.
relationships among different  Streptococci are more likely
phenotypes of the members of to appear in chains when
the family Streptococcaceae grown in broth cultures.
- Currently, there are 17-19  Most members of the genera
genera of catalase-negative, Streptococcus and
gram-positive cocci, including Enterococcus grow in the
Streptococcus, Aerococcus, presence of oxygen but are
Lactococcus, Leuconostoc, and unable to use oxygen for
Pediococcus spp. respiration – considered as
aerotolerant anaerobes
 Carbohydrates are - Organisms in group A
metabolized fermentatively possess the same antigenic C
with lactic acid as the major carbohydrate, organisms in
end product; gas is not group B have the same C
produced carbohydrate, and so on
 Some species are - Some species can
capnophilic, whereas the produce a typespecific
growth of other species is polysaccharide capsule as well
stimulated by increased - These species belong to
CO2, but CO2 is not Lancefield groups A, B, C, D, F,
required. G, and N
 Growth is poor on nutrient - Hemolytic Patterns (Smith’s
media such as TSA, but and Brown’s classification)
more pronounced on media - When lysis of RBCs in the
enriched with blood or agar
serum surrounding the colony is
complete,
Classification Schemes the resulting area is clear – β-
- Four commonly used hemolysis
classification - Partial lysis of the RBCs
schemes are: results in a greenish
 Hemolytic pattern on BAP discoloration of the area
 Physiologic characteristics surrounding the colony and is
 Serologic grouping or typing termed α-hemolysis
of C - When the RBCs
carbohydrate (Lancefield immediately
classification), capsular surrounding the colony are
polysaccharide, or surface unaffected, the bacteria are
protein, described as non-hemolytic
such as the M protein of S. (previously referred to as γ-
pyogenes; hemolysis)
 Biochemical characteristics - Some isolates belonging to
 Lancefield Classification the
Scheme (Cell Wall Structure) viridans group produce what is
- Most streptococci, called wide-zone or α′ hemolysis
except for many of the viridans - Hemolysis
group, have a group or common - These colonies are
C carbohydrate surrounded by a very small
(polysaccharide) – used to zone of no hemolysis and then a
classify an isolate serologically wider zone of β-hemolysis
- This classification - This reaction may be
scheme was developed in the mistaken for β- hemolysis at
1930s by Rebecca Lancefield first glance
- After first recognizing - The use of a dissecting
the antigen, in β-hemolytic microscope or handheld lens
streptococci, Lancefield was reveals the narrow zone of
able to divide the streptococci intact RBCs and the wider zone
into serologic groups, of complete hemolysis
designated by letters
- Physiologic Characteristics o Pyogenic streptococci isolated
(Academic or Bergey’s frequently from humans include
Classification) Streptococcus pyogenes,
- The classification based Streptococcus agalactiae,
on Streptococcus dysgalactiae
physiologic characteristics subsp.
divides equisimilis, and Streptococcus
the streptococcal species into anginosus group (some species
four are
groups: pyogenic streptococci, α-hemolytic or non-hemolytic)
lactococci, enterococci, and - Neither grow in 10 or 45°C,
viridans streptococci grow in 35-37 C Example,
a. Pyogenic streptococci Streptococcus
- Produce pus pyogenes
- Mostly β-hemolytic and - Viridans Group are those that
constitute most of the grow
Lancefield groups at 45°C but not at 10°C.
b. Lactococci are non-hemolytic Example, S.
with mutans, S. sanguis, S. mitis, S.
Lancefield group N antigen and anginosus, S. salivarius
are - Resist classification by
often found in dairy products Lancefield
c. Enterococci comprise species
found as part of the normal Precipitation test
biota - Species that are non-β-
of the human intestine hemolytic (nonpyogenic) non
d. Viridans streptococci pus-forming
- Widely found as normal biota - Enterococcus group that grows
in the upper respiratory tract of at both 10°C and 45°C
humans. - Lactic group grows at 10°C but
- Most strains lack a C not at 45°C, Example
carbohydrate and are not part Streptococcus lactis
of the Lancefield classification;
however, some have A, C, F, G, Clinically Significant
or N antigen Streptococci
- The viridans streptococci are Group A β-hemolytic
α-hemolytic or non-hemolytic streptococci
and are often seen as - Streptococcus pyogenes
opportunistic pathogens - Antigenic Structure
Nevertheless, the terms - cell wall structure similar to
enterococci and viridans that
streptococci remain and are still of other streptococci and gram
used to describe clinical isolates positive bacteria
- Clinically isolated streptococci - The group antigen is unique,
have placing the organism in
historically been separated into: Lancefield group A
- β-hemolytic streptococci or - M protein
pyogenic (pus-forming)
streptococci
- Attached to the peptidoglycan surface together with F proteins
of the cell wall and extends to - Protein F secures the
the cell attachment of the Streptococci
surface in the oral mucosal cells
- Essential for virulence - Antiphagocytic structures
- S. pyogenes colonizes the - Streptococcal pyrogenic
throat exotoxins
and skin on humans, making - Cause a red spreading
these sites the primary sources rash,
of referred to as scarlet fever
transmission - Erythrogenic toxins produced
by
Virulence Factors lysogenic strain of S. pyogenes
- The best-defined virulence when
factor infected with bacteriophage T12
is M protein, encoded by the - Bacteriophage virus infect
genes emm bacteria
- Major virulence factor - The four immunologically
found in cell wall distinct
- No M protein - Avirulent exotoxin types found in
- M protein protect Group A Streptococcus pyogenes are
Streptococci from opsonization SpeA, SpeB, SpeC, and SpeF
by - Streptolysin S
corpulent and subsequent - Non-antigenic, Oxygen
phagocytosis. stable, lyses leukocytes, and
- M protein molecule confers non-immunogenic
resistance to opsonization and - Responsible for
phagocytosis and plays a role in hemolysis in
adherence of the bacterial cell sheep’s blood agar incubated
to aerobically
mucosal cells - Not inhibited by antibody
- The M1 serotype is the most - Streptolysin O
common serotype seen in - Highly antigenic, oxygen
pharyngitis. labile
- More recently, microarray - Lyse RBC, WBC, platelets
analyses of group A streptococci - Responsible for
(GAS) serotype M1 isolates have subsurface
been useful in identifying the hemolysis on BAP incubated
genes associated with anaerobically
pharyngitis - Antibodies measured by
versus invasive disease AntiStreptolysin O
- Lipoteichoic acid and protein F - ASOT – ASO titer for
are tonsilitis or pharyngitis test
adhesion molecules that presence of - Streptolysin O
mediate attachment to host - Streptokinase
epithelial cells - Enzyme produced by
- Lipoteichoic acid affix to Strep. Pyogenes
the - Lyse fibrin clots
protein of the bacterial cell
 - Plasminogen is converted - Invasive infection
into characterized
plasmin, which lyses the fibrin by rapidly progressing
clot inflammation and necrosis of
- Hyaluronic acid capsule the skin, subcutaneous fat, and
- Resistance to phagocytosis fascia
- Encoded by emm genes - Depending on which
- Adhesins organisms
- Help adhere to tissue are cultured, NF may be
- Anti-phagocytic categorized as:
- NF Type 1 - polymicrobial
Clinical Infections infection (aerobic and
1. Bacterial pharyngitis anaerobic bacteria)
- Most common clinical - NF Variant Type 1 – cause
manifestations of GAS by
- Spread through droplets and salt water organism. Ex. Vibrio
close cholerae
contact - NF Type 2 consists of only
- Diagnosis of streptococcal sore GAS
throat relies on a throat culture - NF Type 3 is Gas
or direct antigen detection gangrene or
2. Pyodermal infections Clostridial myonecrosis
- Scarlet fever 4. Streptococcal Toxic Shock
- Strains with temperate Syndrome
bacteriophage T12 produce - A condition in which the entire
streptococcal pyrogenic toxins organ system collapses, leading
- Diffuse red rash on the upper to death
chest, - GAS associated with
trunk and extremities streptococcal TSS produce a
- Desquamation follows after streptococcal pyrogenic
rashes exotoxin, notably SpeA
disappears - major role in the pathogenesis
- Susceptibility Test for Scarlet of this disease functioning as
Fever superantigens leading to
- Dick’s Test - Susceptibility overstimulation of the immune
Test for Scarlet Fever response
- Shaultz-Charlton Reaction - Other virulence factors, such
of Dicks test as
- Blanching SLO and various cell wall
phenomenon antigens, can also contribute to
- Neutralization of toxic shock
erythrogenic toxin when toxin is 5. Poststreptococcal sequelae
injected into skin of patient with - Two serious complications, or
scarlet fever – blanch or remove sequelae, of GAS disease are
skin rash. rheumatic fever (RF, RHD) and
- Used to diagnose when acute glomerulonephritis
rash due to scarlet fever or not (PSGN) -
3. Necrotizing Fasciitis - Entire organ collapse leading
- “flesh-eating disease” to
death 2. Group A Strep. (sensitive)
- Rheumatic fever typically from Group B Strep. (resistant)
follows Streptococcus pyogenes 3. Trimethoprim-
pharyngitis and tonsilitis – most silfamethoxazole (SXT) –
serious result in most serious resistant
and chronic damage of the 4. Sodium Hippurate – negative
heart valve 5. CAMP test – negative
- Acute Rheumatic fever and 6. Lancefield typing – Group A
Chronic sequela – Rheumatic antigen
heart disease Group B, β- hemolytic
- Acute glomerulonephritis Sterptococci
sometimes occurs after a Streptococcus agalactiae
cutaneous or pharyngeal - All strains of Streptococcus
infection, inflammatory damage agalactiae have the group B–
to glomeruli resulting to the specific antigen, an acid-stable
impairment of kidney function polysaccharide located in the
cell wall
Treatment - Colonizes vaginal tract that
- Poststreptococcal sequelae can be
and normal respiratory flora in rare
Acute glomerulonephritis – to few amounts
susceptible to penicillin or
erythromycin Virulence Factors
- Susceptibility test not routinely 1. Capsule
done - The capsule prevents
- For culture, SBA, Incubation phagocytosis but is ineffective
should be at 35°C either in after opsonization
ambient air or under anaerobic - Sialic acid appears to be the
conditions most
- Selective media is SBA significant component of the
containing sulfamethoxazole capsule and a critical virulence
(SMZ) or colistin ang polymyxin determinant
B
Clinical Infections
Colonial Morphology 1. Early-onset infection
- Small - Invasive disease of the
- Gloosy newborn
- Transparent early onset is less than 7 days
- Smooth of
- Large zone of β-hemolytic infant or newborn due o
streptococci congenital transmission from
the mother
Identification and other - Colonization of the vagina and
Biochemical Tests of Strep. rectal area
Pyogenes - Present as pneumonia with
1. PYR test – Positive Bacitracin- meningitis and bacteremia
susceptibility test Sensitive - Screen for β- hemolytic
- Used as presumptive test Sterptococci is recommended
to differentiate for
all pregnant women form 35 – 6. Lancefield typing – Group B
37 antigen
weeks gestation. In adults, the Presumptive identification of
infection affects two types Group A Streptococci are:
of patients - Bacitracin-susceptibility
- First type of patient test susceptible or sensitive
- Young, previously healthy - Group B is a Streptococci
woman who becomes ill after CAMP test – Positive
childbirth or abortion
- Endometritis and wound Other Viridians group o β-
infections are most common hemolytic but (not
- The second type of patient is Group A or Group B) – CAMP
an negative
elderly person with a serious - Group C and G Streptococci
underlying disease or - In the most recent
immunodeficiency classification of β- hemolytic
Susceptibility Test streptococci, isolates from
Suceptibile Penicilin G humans that belong to
Sensitive test are not routinely Lancefield groups
done C and G are subdivided into
Colonial Characteristic large colony and small-colony
- Greyish white mucoid colonies forms
with small zone of β-hemolysis - Large-colony-forming β-
Laboratory Diagnosis hemolytic isolates with group C
- Detection of GBS in pregnant and G antigens belong to the
women is accomplished by subspecies Streptococcus
collecting vaginal and rectal dysgalactiae subsp. equisimilis
material with swabs between 35 - Small-colony-forming β-
and 37 weeks of gestation, hemolytic isolates with group C
incubate 35°C for 18-24 hrs and G antigens belong to the
- Subcultue in Sheep’s blood Streptococcus anginosus
agar group, which are included in the
- Samples should be inoculated viridans streptococci discussed
into later.
selective broth such as:
• Todd-Hewitt broth α-hemolytic Streptococci
contains Streptococcus pneuminiae
colistin and nalidixic acid - Antigenic Structure
▪ C-reactive protein (CRP
Biochemical Test ▪ No Lansfield group
1. PYR test – Negative ▪ Cell wall of S.
2. Bacitracin-susceptibility test- pneumoniae contains an
Resistant antigen, referred to as C
3. Trimethoprim- substance, which is similar to
silfamethoxazole (SXT) – the C carbohydrate of the
resistant various Lancefield groups
4. Sodium Hippurate – Positive ▪ A β-globulin in human
5. CAMP test – Positive serum, called Creactive protein
(CRP), reacts with C substance ▪ Pneumonia resulting
to form a precipitate from S. pneumoniae is not
▪ The amount of CRP usually a
increases during inflammation primary infection but rather a
and infection but Not specific in result of disturbance of the
any part of the body normal defense barriers
- Capsule present 2. Cause secondary bacterial
▪ The characteristic of S. meningitis
pneumoniae that is clearly 3. May also cause otitis media,
associated with virulence is the chorolent sinusitis, peritonitis,
capsular polysaccharide and bacteremia, which often
▪ The capsule is antigenic occurs during the course of a
and can be identified with serious infection
appropriate antisera in the
Neufield test or Neufield- Laboratory Diagnosis
Quellung ▪ Gram-positive cocci in pairs
Reaction) (diplococci)
▪ In the presence of specific ▪ The ends of the cells are
anticapsular serum, the capsule slightly
swells (Quellung reaction) pointed, giving them an oval or
- Exotoxin lancet shape
▪ Can produce hemolysin, ▪ Cocci can occur singly or in
fibrolystin, and leucocidin short chains but most often are
- IgA protease seen as pairs
• will act with IgA antibody ▪ As the culture ages, the Gram
produced by the organism stain
▪ Several toxins are reaction becomes variable,
produced, including a andgramnegative cells are seen
hemolysin, an immunoglobulin ▪ The capsule can be
A protease, demonstrated by using a
neuraminidase, and capsule stain
hyaluronidase ▪ Some isolates require
increased CO2 for growth during
primary isolation
Clinical Infections ▪ They can be anaerobic
1. Pneumococcal pneumonia ▪ Media such as brain-heart
Or Local pneumonia infusion
▪ For an individual to agar, trypticase soy agar with
contract 5%
pneumococcal pneumonia: sheep RBCs, or chocolate agar
▪ The organism must be (CHOC) are necessary for good
present in the nasopharynx growth
▪ The individual must - Procedures used to accomplish
be deficient in the specific this are:
circulating antibody against the ▪ Optochin susceptibility
capsular type of the colonizing test –
strain of S. pneumoniae Streptococcus pneumoniae is
susceptible to optochin
(ethylhydrocuprein HCl)
 ▪ Bile solubility test – referring to the α-hemolysis
Streptococcus pneumoniae is many
soluble to bile species exhibit
▪ Neufield-Quellung - However, β-hemolytic and non
Reaction – for capsule hemolytic species are also
▪ Mauv’s Virulence test classified as viridans
▪ Enzyme such as L- streptococci
alanine - Viridans streptococci are
nuramyle amidase – enzyme fastidious,
cause with some strains requiring CO2
autolysis in Strep. pneuminiae for
▪ Autolysis or “Fitting” of colony growth
produce upon prolong incubation which
causes the sunken center, checkered or Characteristics
mucoid colony - The current classification
▪ Large zone of α-hemolysis assigns
▪ Sunken or pressed center, donut streptococci species in the
shape, nail head – Cultural viridans
characteristic group to one of five groups:
▪ S. mitis group (including
Biochemical Identification S. mitis, S. pneumoniae, S.
- Optochin Disk Susceptibility sanguis, S. oralis)
▪ A filter paper disk ▪ S. mutans group
containing optochin (including S. mutans and S.
(ethylhydrocuprein sobrinus)
hydrochloride or Taxo P) – Taxo ▪ S. salivarius group
P optochin sensitive (including S. salivarius and S.
▪ Most common vestibularis)
presumptive ▪ S. bovis group (including
test used in Streptococcus S. equinus, S. gallolyticus, S.
pneumoniae infantarius, and S. alactolyticus)
▪ Other α-hemolytic ▪ S. anginosus group
Streptococcus are resistant (including S. anginosus, S.
to optochin constellatus, and S.
▪ Best test intermedius)
- Bile susceptibility test - Group D streptococci were
▪ Lysis of S. pneumoniae subdivided into the enterococcal
in the presence of bile salts – and non-enterococcal groups,
Positive Result with the understanding that
α-hemolytic Viridans Group organisms found in the
Streptococci intestinal tract were part of the
- Constituents of the normal enterococcal group
microbiota of the upper - The S. bovis group and the
respiratory tract, the female enterococci possess the group D
genital tract, and the antigen
gastrointestinal tract ▪ Enterococcal
- The term viridans means ▪ bile esculin-positive,
“green,” ▪ grow in a nutrient broth
with 6.5% NaCl
 ▪ placed under genus associated group D antigen in
enterococcus the
▪ non-enterococcal Lancefield classification system
▪ bile esculin-positive, ▪ Normal flora of intestine and
▪ NOT grow in a nutrient mucous membrane but they can
broth with 6.5% NaCl cause exogenous and
▪ remained part of group endogenous disease state of
D streptococci infection

Virulence Factors Laboratory Diagnosis

1. Anginosus group Identification
▪ Polysaccharide capsule ▪Catalase test - weakly
▪ Cytolysin bubbling
▪ Extracellular dextran and ▪ PYR – positive
cell ▪ Bile esculin – positive
surface–associated proteins ▪ Grow in 6.5% NaCl
(adhesins) are implicated in ▪ In contrast to streptococci,
adherence and colonization of enterococci have the ability to
these grow under extreme conditions
organisms in endocarditis – for example, in the
2. Groups C and G streptococci presence of bile or 6.5% NaCl or
▪ Possess M proteins with at
similarity to GAS 45°C or alkaline pH
▪ Some of them also ▪ Resistant to multiple
produce strain of antibiotic
extracellular enzymes such as ▪ Vancomycin Resistant Strain
SLO, of
hyaluronidase, and DNase Enterococcus spp. Exist
▪ Perform only when isolate is
Laboratory Diagnosis clinically significant
▪ PYR -negative (all member) ▪ Other urologic manipulation
▪ leucine aminopeptidase (LAP)- can cause UTI and bacteremia
positive
▪ Optochin sensitivity – Media
resistant to ▪ Trypticase soy agar with 5%
optochin sheep blood and grow well at
▪ Bile solubility tests – bile 35°C in the presence of CO2
insoluble ▪ selective media containing
▪ Bile esculin the following should be used for
▪ Growth in 6.5% NaCl isolation of enterococci:
▪ Bile esculin azide
Enterococcus Group ▪ Colistin-nalidixic acid
▪ The commonly identified ▪ Phenylethyl alcohol
species in ▪ Chromogenic substrates
clinical specimens are E. ▪ Cephalexin-aztreonam-
faecalis and E. faecium arabinose agar
▪ All species produce the cell ▪ Enterococcus spp. are
wall identified based on their:
▪ Ability to produce acid in
carbohydrate broth non-hemolytic group D
▪ Ability to hydrolyze streptococci
arginine ▪ The PYR test detects the
▪ Tolerance of 0.04% activity of L-pyrrolidonyl
tellurite arylamidase, also called
▪ Utilization of pyruvate pyrrolidonyl aminopeptidase
▪ Ability to produce acid ▪ The PYR test is more
from methyl-α- D- specific
glucopyranoside for S. pyogenes than bacitracin
▪ Growth around 100-μg ▪ Streptococcus pyogenes is
efrotomycin acid disk the
▪ Motility only species of Streptococcus
that is PYR-positive
Identification of
Streptococcus and Christie, Atkins, Munch-Petersen
Enterococcus (CAMP) Test
1. Hemolytic pattern on BAP ▪ Used for presumptive
2. Physiologic characteristics identification of GBS that
3. Serologic grouping or typing differentiate Streptococcus
of C agalactiae from other β-
carbohydrate (Lancefield hemolytic streptovocci
classification), capsular ▪ The CAMP test can beS
polysaccharide, or surface performed three ways:
protein, 1. use of a β-lysin
such as the M protein of S. producing strain of
pyogenes; Staphylococcus aureus
4. Classification Scheme or 2. use of a disk
biochemical test impregnated with the β-
Biochemical Identification lysin
▪ Bacitracin Susceptibility 3. rapid CAMP test (or spot
- helpful in screening for CAMP test)
Group AS ▪ The first two methods take
- pattern shown by S. advantage of the enhanced
pyogenes hemolysis occurring when the
(group A) is susceptibility to β-lysin and the hemolysin
bacitracin and resistance to produced by GBS are in contact
SMZ ▪ result is a characteristic
- pattern shown by S. arrowhead-shaped hemolysis
agalactiae pattern
(group B) is resistance to both
bacitracin and SMZ Hippurate Hydrolysis
- use blood agar ▪ test for differentiating
Streptococcus agalactiae from
Pyrrolidonyl-α-Naphthylamide other β-hemolytic streptococci
(PYR) Hydrolysis ▪ Streptococcus agalactiae
▪ presumptive identification possess the enzyme hippuricase
of (also called hippurate
the β-hemolytic GAS and the hydrolase), which hydrolyzes
sodium hippurate to form identification schemes for non-
sodium benzoate and glycine hemolytic, catalasenegative,
to give POSITIVE result gram-positive cocci
▪ The bile esculin test is a
Voges-Proskauer (VP) Test two-step test detecting growth
▪ distinguish the small- of bacteria in the presence of
colony-forming β-hemolytic 40% bile and the ability to
anginosus group containing hydrolyze esculin
groups A or C antigens from ▪ Group D streptococci and
large-colony–forming pyogenic Enterococcus spp. are positive
strains with the same antigens ▪ Organisms positive for
bile esculin are separated into
Leucine Aminopeptidase (LAP) group D
▪ LAP is a peptidase that streptococci or Enterococcus by
hydrolyzes peptide bonds the
adjacent to a free amino group salt-tolerance test
▪ LAP reacts most quickly
with leucine, it is called leucine Optochin Susceptibility
aminopeptidase ▪ A filter paper disk
▪ LAP test is often used with containing
the PYR test and is most helpful optochin (ethylhydrocuprein
in hydrochloride or Taxo P) at 5
differentiating Aerococcus and μg/mL
Leuconostoc spp. from other or less (excess of 5 μg/mL
gram inhibits
positive cocci viridans streptococci as well)
▪ Streptococcus, ▪ Presumptive test for
Enterococcus, and Pediococcus Streptococcus
spp. are LAP-positive pseudopneumoniae
▪ Aerococcus and ▪ Isolates producing smaller
Leuconostoc spp. are LAP- zones
negative should be tested for bile
solubility to
β-D-glucuronidase Test confirm their identity
▪ Test detects the action of ▪ A zone of inhibition greater
β-D than 14 mm with a 6-mm disk
glucuronidase, an enzyme found or a zone of inhibition greater
in than 16 mm with a 10-mm disk
isolates of large-colony-forming is considered
β- susceptible and a presumptive
hemolytic groups C and G identification of S. pneumoniae
streptococci but not in the small
colony-forming β-hemolytic Bile Solubility Test
anginosus group ▪ This test correlates well
with
Bile Esculin and Salt Tolerance optochin susceptibility
▪ The bile esculin hydrolysis
test and the salt-tolerance test Immunoassays
have been mainstays in
 ▪ ID of streptococci,
particularly GAS, can be
accomplished by detection of
group-specific antigen from
either isolated colonies
Automated:
▪ ELISA Enzyme linked
Immunoassay
▪ Nucleic acid probe test