UNIT VI: STREPTOCOCCI

STREPTOCOCCACEAE
FAMILY ➢ Part as normal flora of the human body; however, gaining access to normally
sterile sites makes them cause life-threatening infections

➢ Gram-positive, spherical-shaped, arranged in chains or pairs

➢ Catalase-negative
➢ Non-motile (no flagella)
➢ Non-spore former
➢ Young cultures of some species can produce capsules
○ e.g., Streptococcus pneumoniae
➢ Oxidase-negative
➢ Do not produce gas
➢ Ferment carbohydrates
➢ Growth is more pronounced on a blood agar plate (BAP) but poor on a
nutrient agar plate (NAP)
○ Growth is enhanced by blood, serum, or mucus present in an agar
plate
➢ Facultative anaerobe
GENERAL CHARACTERISTICS ○ Can multiply either in the presence or absence of oxygen; can use
oxygen as the final electron acceptor in cellular respiration
○ Can acquire energy from fermentation
○ Peptostreptococcus – obligate anaerobe
■ Unable to multiply in the presence of oxygen
■ Require incubation in anaerobic environments (e.g., candle
jar, gas pack)
➢ Some species are capnophilic
○ Capnophilic: require an increased amount of carbon dioxide for
growth
➢ Small and somewhat transparent
○ Grayish, pinpoint, translucent, slightly opaque colonies
➢ Mucoid consistency
➢ Possess different hemolytic patterns (α, β, γ)

BROWN’S CLASSIFICATION OF HEMOLYSIS (TYPE AND PATTERN OF

HEMOLYSIS)
CLASSIFICATIONS OF STREPTOCOCCI ➢ Colonial morphology and hemolytic patterns on agar
 SHERMAN’S CLASSIFICATION
➢ Physiologic divisions
○ Pyogenic (pus-forming)
○ Viridans
○ Lactic
○ Enterococcus

ANTIGENIC PROPERTIES
➢ Lancefield antigen grouping (serological specificity of the cell wall
carbohydrate group antigen)

BERGEY’S CLASSIFICATION
➢ Three genera
○ Streptococcus
○ Enterococcus
○ Lactococcus

BAP is traditionally 5% defibrinated sheep blood in trypticase soy agar.

ALPHA (α)
➢ Incomplete or partial hemolysis, resulting from hydrogen peroxide production
by the organism
○ Release of hemoglobin from destroyed RBCs
➢ Green or brown zone
➢ Production of streptolysins S and O
➢ Require oxygen during incubation
○ Become non-hemolytic in anaerobic environments
BROWN’S CLASSIFICATION OF HEMOLYSIS ➢ e.g., S. pneumoniae and Viridans group
J.H. Brown*, 1919
BETA (β)
➢ Complete hemolysis/disruption of red blood cells, with clearing
➢ Colonies are surrounded by haloes
➢ e.g., S. pyogenes
○ Naturally pathogenic Streptococcus

GAMMA/NONHEMOLYTIC (γ)
➢ Lack of hemolysis
➢ No color change
➢ e.g., Group D Streptococci (Enterococcus)
 ALPHA PRIME
➢ A small zone of α-hemolysis surrounds the colony
➢ A small zone of β-hemolysis surrounds the zone of α-hemolysis

➢ Based on hemolytic reactions, carbohydrate antigens, and phenotype tests

○ Pyogenic
■ β-hemolytic strains A, B, C, D, E, F, and G
■ Neither grow on 10 or 45 degrees Celsius, only 37 degrees
Celsius
○ Viridans
■ Not ß-hemolytic and salt-tolerant; able to grow at high pH
■ Will grow both at 45 and 37 degrees Celsius
■ Normal flora in the human upper respiratory tract
SHERMAN’S CLASSIFICATION ○ Lactococci
Sherman, 1973
■ Not clinically significant but associated with the dairy
industry
■ Will grow in 10 and 37 degrees Celsius
■ Often found in dairy products
■ e.g., S. lactis (causes normal coagulation in the souring of
milk)
○ Enterococcus
■ Salt-tolerant; can grow at high pH and within 10–45 degrees
Celsius

➢ β-hemolytic strains classified based on specific carbohydrate group antigen

by immunochemical methods
LANCEFIELD ANTIGEN GROUP ○ CHO in the cell wall antigen is extracted, and the organism is
Rebecca Lancefield, 1933 serotyped with a type-specific antiserum
➢ First five groups: A, B, C, D, and E
○ Other groups: F and G

CLINICALLY SIGNIFICANT STREPTOCOCCI

➢ “Fever-producing bacteria,” “flesh-eating bacteria”

➢ β-hemolytic strep
Streptococcus pyogenes
➢ Almost always clinically significant
GROUP A STREPTOCOCCUS (GAS) ➢ Group-specific antigenic substance: A
➢ May be found on the skin, upper respiratory tract, and pharyngeal mucosa
 (but not normal flora)
➢ Causes the following:
○ Bacterial pharyngitis (strep throat)
■ 80 different strains of S. pyogenes
○ Skin infections
○ Complications, such as rheumatic heart disease and acute
glomerulonephritis

MODES OF TRANSMISSION
➢ Direct person-to-person contact
➢ Indirectly in aerosol droplets from coughing or sneezing

ANTIGENIC STRUCTURE/VIRULENCE FACTORS

➢ Lancefield group A
➢ M (emm protein): Cell surface antigen
○ Contains hyaluronic acid
○ Surface protein found in encapsulated strains of S. pyogenes
○ Hairlike projections of the streptococcal cell wall, associated with
virulence (major virulence factor)
○ Makes S. pyogenes appear smooth, glossy, round, and mucoid
○ Over 200 types
○ Holds antiphagocytic properties and permits the bacterium to survive
within the host
○ Attached to the peptidoglycan of the cell wall and extends to the cell
surface
➢ Protein F (fibronectin-binding protein)
○ Mediates epithelial cell attachment
➢ Lipoteichoic acid
○ Bacterial adherence to the respiratory epithelium
○ Secures the attachment of streptococci to the oral mucosal cells
together with the M protein
➢ Hyaluronic acid capsule
○ Weakly immunogenic
○ Prevents opsonized phagocytosis by neutrophils and macrophages
○ Allows the bacterium to mask its antigens and remain unrecognized
by its host
➢ Hemolysins
○ Lyse the membranes of RBCs and damage other cell types
○ Streptolysin O (exclusive to S. pyogenes)
 ■Oxygen-labile (active in reduced form)
■Highly antigenic (can produce antibodies)
■Also causes lysis of WBCs, platelets, and tissue cells
■Induces antibody response
■Responsible for subsurface hemolysis on BAP incubated
anaerobically
● Stab or cut down agar plate to force organisms into
growing within reduced oxygen content
■ Antistreptolysin O (ASO) test: Detects recent infection with
S. pyogenes
○ Streptolysin S
■ Oxygen-stable
■ Non-antigenic
■ Responsible for surface hemolysis on BAP incubated
aerobically
■ Also causes WBC lysis
■ Effect is elaborated in the presence of serum
■ Can determine past S. pyogenes infections
● Most S. pyogenes strains produce both
streptolysins; only 10% produce either
➢ Toxins and enzymes
○ Deoxyribonuclease (DNase)
■ 4 types: A, B, C, and D (antigenic enzymes)
■ Lowers viscosity of exudates, giving pathogens more
mobility
■ Antibodies to these enzymes can be detected following
infections
○ Streptokinase (fibrinolysin)
■ Causes lysis of fibrin clots
■ A protein that binds to plasminogen and activates the
production of plasmin
● Plasmin: An active proteolytic enzyme that digests
fibrin and other proteins, allowing bacteria to escape
from the blood clot
■ Activates a host-blood factor that dissolves fibrin clots
○ Hyaluronidase
■ Solubilizes the ground substance of mammalian tissue
■ Spreading factor (aids in spreading the infective organism)
■ To separate the tissue and spread the organism
 ○ Streptococcal pyrogenic exotoxins (Spes)
■ Formerly known as erythrogenic toxins
■ Cause red spreading rash (Scarlet fever)
■ 3 immunologically distinct exotoxin types: SpeA, SpeB,
SpeC
● Toxins function as superantigens

INFECTIONS AND DISEASES

➢ Pharyngitis or tonsillitis (strep throat)
○ Spread by droplets and close contact
○ Diagnosis: throat culture or direct antigen detection
○ Incubation period: 1–4 days
○ Most common infection
○ Caused by adherence to pharyngeal epithelium by means of
lipoteichoic acid-covered surface pili and hyaluronic acid capsule
○ S/S: sore throat, malaise, fever and headache, inflammation of
tonsils and pharynx, swollen cervical lymph nodes
■ Signs and symptoms subside within 3–5 days, unless
complications occur, such as peritonsillar abscesses
➢ Scarlet fever (Scarlatina)
○ Communicable, spread via inhalation of infectious respiratory
droplets
○ Caused by temperate bacteriophage T12 that produces Spe
○ Immunity to toxin is demonstrated by the Dick test
■ Injection of toxin
■ If local antibodies are absent, local erythema occurs
○ Cardinal signs: diffuse red rash on upper chest and spread through
the trunk and the rest of the body
○ Tongue becomes strawberry red
○ Rash disappears in 5–7 days and is followed by desquamation
➢ Skin or pyodermal infections
○ Impetigo
■ Localized lesion
■ Pyoderma: Any confined, pus-producing lesion that usually
occurs on the exposed skin of the face, arms, and legs
(similar to impetigo)
○ Cellulitis
■ A diffuse spreading infection of subcutaneous skin tissue
characterized by a defined area of redness (erythema) and
 accumulation of fluid (edema)
■ Compared to erysipelas, the lesion is not raised, and the line
between the involved and uninvolved tissue is not distinct
■ Progresses into gangrene
○ Erysipelas
■ Usually occurs in faces of children
■ Acute spreading lesion that is erythematous with
demarcated but irregular edges
■ An acute infection and inflammation of the dermal layer of
the skin characterized by painful reddish patches that
enlarge and thicken with a sharply defined edge
○ Wound infection
○ Necrotizing fasciitis, aka “flesh-eating disease”
■ Other names: suppurative fasciitis, hospital gangrene,
galloping gangrene, necrotizing erysipelas, flesh-eating
bacteria syndrome
■ Caused by flesh-eating strep
■ An invasive infection characterized by a rapidly progressing
inflammation and necrosis of the skin, subcutaneous fat, and
fascia
■ A skin infection caused by Group A streptococci strains may
also lead to necrotizing fasciitis
■ May be characterized into type 1 (polymicrobial infection,
either aerobic or anaerobic), type 2 (caused by GAS), or
type 3 (gas gangrene or clostridial myonecrosis)
○ Arthritis
➢ Septic infections
○ Streptococcal toxic shock syndrome (TSS)
■ Group A strep can spread from initial site of infection (HIV,
cancer, PD, HD, diabetes mellitus) which leads to
bacteremia and severe multi-system infections
■ Entire organ system shuts down, leading to death
■ The initial strep infections include pharyngitis, cellulitis, and
wound infections
■ SpeA plays a major role in the pathogenesis of this disease
■ Clinically overlaps with scarlet fever
➢ Post-streptococcal sequelae
○ Rheumatic fever (most serious post-strep sequelae)
■ Characterized by fever, inflammation of the heart, joints, and
 blood vessels
■ Complication of pharyngitis
■ S/S: heart murmur from narrowed mitral valves, erythema
marginatum (non-itchy rash), painful joints, fever of
38.2–38.8 degrees Celsius
■ Potential risk factors: repeated strep infections, contracting
scarlet fever, skipping antibiotics for strep or scarlet fever
treatment, not practicing proper hand hygiene
○ Acute glomerulonephritis
■ Inflammatory disease of the renal glomeruli, resulting from
the deposition of antigen-antibody complexes
■ Likewise a complication of pharyngitis
■ Causes hypertension due to blockage

TREATMENT
➢ Penicillin
➢ Erythromycin
➢ Cephalosporin
➢ Bacitracin
➢ Surgical removal of non-viable tissues and infecting bacteria (in necrotizing
fasciitis)

Antigenic structure: Group B-specific antigen

➢ Part of the normal flora of female genital tract and lower GIT
➢ Nosocomially transmitted by unwashed hands of the mother or a healthcare
personnel to the newborn infant
➢ Causes infections of fetuses
○ It is recommended that all pregnant women be screened for group B
streptococci at 35–37 weeks of gestation
Streptococcus agalactiae
➢ Stages of neonatal infection
GROUP B STREPTOCOCCI ○ Colonization of pregnant mother
○ Ascending placental and uterine infection
○ Pneumonia and lung injury
○ Bacteremia and sepsis
○ Blood-brain barrier penetration and meningitis
➢ Molecular and cellular pathogenesis
○ Extracellular matrix binding
○ Direct cellular injury
 ○ Resist intracellular killing
○ Brain endothelial invasion
○ Epithelial cell invasion
○ Resist phagocytosis
○ Inflammatory activation
○ CNS neutrophil recruitment

DEFINITION
➢ Gram-positive cocci in chains
○ 1–2 mm colonies (in diameter)
➢ Catalase negative
➢ Oxidase negative
➢ Non-spore former
➢ Non-motile
➢ The only species positive for CAMP test and can hydrolyze sodium hippurate

DISEASES AND INFECTIONS

➢ Pneumonia
➢ Meningitis
➢ Neonatal sepsis
➢ Postpartum infection
➢ Osteomyelitis
➢ Urinary tract infection
➢ Endocarditis

VIRULENCE FACTORS
➢ Capsule
○ Sialic acid
➢ Hemolysin
➢ CAMP factor
○ Unique/exclusive to GBS
➢ Neuraminidase
➢ DNase
➢ Hyaluronidase
➢ Protease

➢ Recovered from the upper respiratory tract, vagina, and human skin
GROUP C AND G STREP ➢ Possess M protein (like group A streptococci)
○ Mimic GAS in BAP
 ➢ Group C streptococci: main source of streptokinase, and an animal pathogen
➢ Species: S. dysagalactiae sp. equisimilis, S. equi sp. zooepidemicus
○ S. dysagalactiae sp. equisimilis: upper RT, vagina, and human skin;
spectrum of infections resembles S. pyogenes; antigenic groups C,
G, L, and A
○ S. equi sp. zooepidemicus: animal pathogen, rarely human;
associated with acute glomerulonephritis and rheumatic fever
following infections
➢ Cause bacterial pharyngitis, sinusitis, bacteremia, and endocarditis
➢ Differentiated from GAS through immunochemical methods
○ Determining carbohydrate group antigen
➢ Forms:
○ Large colony-forming isolates with group A, C, and G antigens are
classified with pyogenic streptococci
○ Large colony-forming β-hemolytic isolates with groups C and G
antigens belong to the S. dysagalactiae sp. equisimilis
○ Small colony-forming β-hemolytic isolates with groups C and G
antigens belong to S. anginosus group (included in the viridans
streptococci)

➢ Second most clinically important Streptococci

➢ Aka diplococcus/pneumococcus
➢ Normal flora (20–50%) of the URT
➢ Common isolate
➢ May be pathogenic, particularly in preschool children
➢ Causes lobar pneumonia and bacterial meningitis in adults
○ Lobar pneumonia: most common bacterial pneumonia in elderly

VIRULENCE FACTORS
Streptococcus pneumoniae ➢ C-substance
○ Antigen on its cell wall
○ Reacts with the β-globulin human serum (C-reactive protein)
■ Positive (+) result: precipitation
○ Similar to C carbohydrate in Lancefield group
➢ Capsule (capsular polysaccharide)
○ The most important virulence factor
○ There are >80 different types and antibodies against the capsule
○ Type-specific and protective
○ Differs from Group C streptococci, such that GCS are not
 encapsulated
➢ Toxins
○ Hemolysin, immunoglobulin A protease, neuraminidase,
hyaluronidase

CLINICAL INFECTIONS
➢ Bacterial meningitis
○ Usually follows other Streptococcus pneumoniae infections, such as
otitis media or pneumonia

TREATMENT
➢ Penicillin
➢ Cephalosporin
➢ Erythromycin
➢ Chloramphenicol

➢ Oropharyngeal commensals that are regarded as opportunistic pathogens

➢ Produce green strains
➢ α–hemolytic (α’ streptococci)
○ Some β-hemolytic, some non-hemolytic
➢ Fastidious and capnophilic
➢ Pathogenic in low virulence
➢ Lack Lancefield group antigens and do not fall in the criteria of S.
pneumoniae
➢ Normal microbiota of the URT, female genital tract, and GIT
➢ More than 30 species
➢ 5 groups:
Viridans streptococci ○ S. mitis group
○ S. mutans group
○ S. salivarius group
○ S. bovis (equinis) group
○ S. anginosus group
➢ Transient bacteria is associated with endocarditis
➢ May cause fulminant cardiovascular collapse or meningitis
➢ May infect the bloodstream, oral cavity (e.g., gingivitis, dental caries/cavities)
➢ Other diseases: meningitis, abscesses, osteomyelitis, and empyema

S. mitis group S. mutans group S. salivarius group

 ➢ S. mitis
➢ S. sanguinis
➢ S. parasanguinis
➢ S. salivarius
➢ S. gordonii ➢ S. mutans
➢ S. vestibularis
➢ S. cristatus ➢ S. sobrinus
➢ S. thermophilus
➢ S. infantis
➢ S. oralis
➢ S. peroris

S. equinis S. anginosus

➢ S. anginosus
➢ S. equinis
➢ S. constellatus
➢ S. gallolyticus
➢ S. intermedius
➢ S. infantarius
*Lancefield group A, C, F, G, or
➢ S. alactolyticus
non-antigenic/non-groupable at all in some
*Contain Group A antigens
instances

➢ A commensal of the normal oral and GI microbiota

Viridans streptococci
➢ Associated with abscess formation in pharynx, brain, and peritoneal cavity
S. anginosus ➢ Isolated from bacteremic patients with invasive pyogenic infections with
tendency to form abscess

➢ Most commonly isolated group

Viridans streptococci
➢ Isolated from the oral cavity
S. mutans ➢ Primary contributor to dental caries
➢ Most common member associated with bacteremia

➢ Previously known as Group D streptococci

➢ Commensals of the intestinal tract of humans and animals
➢ Most are non-hemolytic or α-hemolytic; some β-hemolytic
➢ Most commonly identified species:
○ E. faecalis and E. faecium
➢ Other species:
Enterococcus ○ E. durans
○ E. casseliflavus
○ E. gallinarum
○ E. raffinosus
➢ Possess the D substance/antigen/carbohydrate (Lancefield classification)
➢ Nonpathogenic but frequently cause nosocomial infections
➢ Resistant to multiple antimicrobial agents
 VIRULENCE FACTORS (E. faecalis)
➢ Extracellular surface protein
➢ Extracellular serine protease
➢ Gelatinase
➢ Cytolysin

CLINICAL IMPLICATIONS
➢ Nosocomial infections
➢ UTI
➢ Bacteremia

LABORATORY DIAGNOSIS OF STREPTOCOCCUS

I. Hemolytic pattern
➢ Group A: β-hemolytic
○ S. pyogenes: suppurative diseases (pharyngitis, cellulitis),
and nonsuppurative (rheumatic fever and
glomerulonephritis)
➢ Group B: β-hemolytic
○ S. agalactiae: neonatal meningitis and sepsis
➢ Group C: β-hemolytic (S. equisimilis)
➢ Group D
○ β-hemolytic on rabbit blood agar
○ α- or γ- hemolytic in sheep blood agar
➢ Group F: β-hemolytic (S. anginosus)
CLASSIFICATION SCHEME ➢ Group G: β-hemolytic
○ Part of the GIT, vaginal and oropharyngeal and skin flora
II. Physiologic characteristics (academic classification)
➢ Pyogenic, Viridans, Lactis
III. Serologic grouping or typing of C carbohydrate (Lancefield classification)
IV. Biochemical identification
A. Bacitracin susceptibility test
B. CAMP (Christie, Atkins, Munch-Petersen)
C. Hippurate hydrolysis
D. PYR hydrolysis (L-pyrrolidonyl-β-naphthylamide)
E. Leucine aminopeptidase (LAP)
➢ Often used with PYR
➢ Most helpful in differentiating Aerococcus and Leuconostoc
 (LAP-negative) from Gram-positive cocci
➢ Streptococci, Enterococcus, Micrococcus: LAP-positive
F. Voges-Proskauer (VP)
➢ For Enterococcaceae
➢ Groups A (+) and C (+) Streptococci
G. β-D-glucuronidase
➢ Action of the said enzyme in isolates of large colony-forming
unit β-hemolytic groups C and G
➢ Cannot be found in small colony-forming unit β-hemolytic S.
anginosus group
H. Bile solubility

I. Direct Gram-stained smears (microscopic examination)

➢ Gram-positive in chains or pairs
II. Culture media (BAP)
A. Hemolytic pattern
B. Biochemical identification
III. Non-cultural identification (serologic testing)
➢ Determination of serogroups through capillary precipitin used by
Lancefield based on C substance in cell wall
➢ Determination of serotypes—Group A Streptococci are divided into
>80 different serotypes based on the type of M protein that they
express
➢ Quellung Test: used to identify the >84 different types of
Streptococcus pneumoniae based on difference in the type of
IDENTIFICATION OF STREPTOCOCCI specific soluble substances (SSS) in their capsule
○ 𝑆𝑢𝑠𝑝𝑒𝑛𝑠𝑖𝑜𝑛 + 𝑎𝑛𝑡𝑖𝑠𝑒𝑟𝑢𝑚 + 𝑚𝑒𝑡ℎ𝑦𝑙𝑒𝑛𝑒 𝑏𝑙𝑢𝑒 = 𝑠𝑤𝑒𝑙𝑙𝑖𝑛𝑔
IV. Susceptibility testing
➢ Bacitracin disc susceptibility test (0.04 units)
○ Presumptive identification of Group A (β-hemolytic)
■ GAS: Susceptible
■ GBS: Resistant
○ Most helpful method in identifying GAS (particularly in throat
specimens)
➢ Trimethoprim sulfamethoxazole (SXT) disc susceptibility test
○ Used to distinguish groups A and B from other β-hemolytic
species
■ GAS and GBS are resistant
➢ CAMP test
 ○ Christie, Atkins, Munch-Petersen
○ Presumptive identification of GBS
○ Hemolytic activity of S. aureus (β) is enhanced by an
extracellular protein produced by GBS
○ Positive (+) result: arrowhead
○ Other method: rapid/spot CAMP test
■ Drop of extracted β-lysin
➢ Hydrolysis of sodium hippurate
○ Identification of GBS
○ A 2-hour test that detects hippuricase (hippurate hydrolase):
enzyme possessed by GBS
○ Hippurate = glycine + sodium benzoate
○ Reagents:
■ Ferric chloride = heavy precipitate (within 10
minutes)
■ Ninhydrin reagent = deep blue color (glycine)
➢ Hydrolysis of L-pyrrolidonyl-β-naphthylamide (PYR)
○ Identification of groups A and D (both positive)
○ Detects L-pyrrolidonyl-arylamidase (pyrrolidonyl
aminopeptidase)
𝑃𝑌𝑅 𝑖𝑚𝑝𝑟𝑒𝑔𝑛𝑎𝑡𝑒𝑑 𝑑𝑖𝑠𝑐𝑠 = 𝐿 − 𝑝𝑦𝑟𝑟𝑜𝑙𝑖𝑑𝑜𝑛𝑒 𝑎𝑛𝑑 β − 𝑛𝑎𝑝ℎ𝑡ℎ𝑦𝑙𝑎𝑚𝑖𝑑𝑒
+ 𝑝 − 𝑑𝑖𝑚𝑒𝑡ℎ𝑦𝑙𝑎𝑚𝑖𝑛𝑜𝑐𝑖𝑛𝑛𝑎𝑚𝑎𝑙𝑑𝑒ℎ𝑦𝑑𝑒 (𝑟𝑒𝑑 𝑐𝑜𝑙𝑜𝑟)
○ Positive result in Enterococcus, Aerococcus, and Gemellas
➢ Bile esculin test
○ Two-step test
■ Hydrolysis of esculin in the presence of 40% bile
■ Identification of growth in Enterococcus and Group
D (non-hemolytic, Gram-positive, catalase-negative
cocci)
○ Use of bile esculin medium
■ Positive (+) result: blackening of agar
➢ Salt tolerance test (6.5% NaCl broth)
○ Test for the ability of an organism to grow in 6.5% NaCl
broth
○ (+) Enterococcus, Aerococcus, some Pediococcus and
Leuconostoc (24 hours incubation)
○ (-) Group D Streptococci
➢ Susceptibility to ethyl hydrocupreine hydrochloride (optochin test)
○ Inhibits the growth of S. pneumoniae
 ○ Used to differentiate S. pneumoniae from other α-hemolytic
Streptococci
○ Basis: ZOI
■ >14 mm around 6-mm disc or >16 mm around
10-mm disc: (+)
○ S. mitis: resistant to optochin
➢ Bile solubility test
○ Identification of S. pneumoniae
○ Under the influence of bile salts or detergent
○ Sodium deoxycholate and sodium taurocholate can lyse S.
pneumoniae
■ Presence of enzyme, autocatalytic amidase
○ (+) results:
■ Clearing of 10% sodium deoxycholate
■ Disappearance of colonies
○ Confirmatory test for Streptococci with ZOI <14 mm

➢ Other Gram-positive cocci (Streptococci) are catalase-negative, while

Staphylococci and Micrococci are catalase-positive
➢ Other cocci (Neisseria) are Gram-negative, but since Streptococci quickly
DIFFERENTIATION FROM OTHER GENERA lose their Gram-positive characteristics as they age, they may be confused
with Neisseria
➢ Neisseria spp. are oxidase-positive and Streptococcus spp. are
oxidase-negative

➢ Respiratory tract
○ α-hemolytic are likely to be viridans group Streptococci
○ Streptococcus pneumoniae (pathogen)
➢ GI tract
○ α-hemolytic is probably Enterococcus
○ β-hemolytic Streptococcus are likely to be Streptococcus pyogenes
DIFFERENTIATION WITHIN THE GENUS
(Group A)
(SOURCE OF SPECIMEN) ➢ Genitourinary tract
○ β-hemolytic Streptococci are likely to be Streptococcus agalactiae
(Group B)

Differential diagnosis: certain strep are more commonly associated with specific
clinical syndromes (i.e., Group A with sore throat and Group B with meningitis)
 KIT SYSTEM
➢ API rapid STREP
○ One large strip with wells
OTHER TESTS ➢ RapID STR
➢ Vitek Gram-positive identification (most used in the laboratory)
➢ API 2005
➢ Minitek Gram-positive test