Giáo trình Streptococci

Nội dung text: Giáo trình Streptococci

1. Streptococci • Characters of Streptococci • Cầu khuẩn, Gram + • Đường kính 1µm • Chuỗi • Thương có vỏ bọc • Không di động • Không sinh nha bào Nuôi cấy • Yếm khí tùy tiện • Khó tính • Catalase negative (Staphylococci are catalase positive)
2. Classification of Streptococci • Streptococci can be classified according to: • Oxygen requirements (Cần oxy) • Anaerobic (Peptostreptococcus) • Aerobic or facultative anaerobic (Streptococcus) • Serology (Lanciefield Classification) • Hemolysis on Blood Agar (BA)
3. Serology: Lanciefield Classification Streptococci Lanciefield classification Group A Group B Group C Group D Other groups S. pyogenes S. agalactiae S. equisimitis Enterococcus (E-U) • Streptococci classified into many groups from A-K & H-V • One or more species per group • Classification based on C- carbohydrate antigen of cell wall • Groupable streptococci • A, B and D (more frequent) • C, G and F (Less frequent) • Non-groupable streptococci • S. pneumoniae (pneumonia) • viridans streptococci • e.g. S. mutans • Causing dental carries
4. Classification of Streptococci Based on Hemolysis on Blood Agar Hemolysis on BA – -hemolysis Partial hemolysis Green discoloration around the colonies e.g. non-groupable streptococci (S. pneumoniae & S. viridans) – -hemolysis Complete hemolysis Clear zone of hemolysis around the colonies e.g. Group A & B (S. pyogenes & S. agalactiae) – -hemolysis No lysis e.g. Group D (Enterococcus spp) Streptococci -hemolysis -hemolysis -hemolysis
5. Hemolysis on Blood agar -hemolysis -hemolysis -hemolysis
6. Group A streptococci • Include only S. pyogenes • Group A streptococcal infections affect all ages peak incidence at 5-15 years of age (lứa tuổi mắc ở người 5-15 tuổi) • 90% of cases of pharyngitis (viêm họng)
7. Pathogenesis and Virulence Factors • Structural components • M protein M, which interferes with opsonization and lysis of the bacteria • Lipoteichoic acid & F protein adhesion • Hyaluronic acid capsule, which acts to camouflage the bacteria • Enzymes • Streptokinases • Deoxynucleases facilitate the spread of streptococci through tissues • C5a peptidase • Pyrogenic toxins that stimulate macrophages and helper T cells to release cytokines • Streptolysins • Streptolysin O lyse red blood cells, white blood cells, and platelets • Streptolysin S
8. Opsonization
9. Disease caused by S. pyogenes • Suppurative (sự sinh mủ) • Non-Invasive • Pharyngitis (“strep throat”)-inflammation (viêm họng) of the pharynx • Skin infection, Impetigo (bệnh chốc lở) • Invasive • Scarlet fever-rash (sốt ban đỏ) that begins on the chest and spreads across the body • Pyoderma-confined, pus-producing lesion (viêm da mủ) that usually occurs on the face, arms, or legs • Necrotizing fasciitis-toxin production (độc tố gây viêm cơ da hoại tử) destroys tissues and eventually muscle and fat tissue • Non Suppurative • Rheumatic fever (sốt thấp khớp): Life threatening inflammatory disease that leads to damage of heart valves muscle • Glomerulonephritits (viêm tiểu cầu thận) • Immune complex disease of kidney • inflammation of the glomeruli and nephrons which obstruct blood flow through the kidneys
10. Differentiation between -hemolytic streptococci • The following tests can be used to differentiate between -hemolytic streptococci • Lanciefield Classification • Bacitracin susceptibility Test • Specific for S. pyogenes (Group A) • CAMP test • Specific for S. agalactiae (Group B)
11. Bacitracin sensitivity • Principle: • Bacitracin test is used for presumptive identification of group A • To distinguish between S. pyogenes (susceptible to B) & non group A such as S. agalactiae (Resistant to B) • Bacitracin will inhibit the growth of gp A Strep. pyogenes giving zone of inhibition around the disk • Procedure: • Inoculate BAP with heavy suspension of tested organism • Bacitracin disk (0.04 U) is applied to inoculated BAP • After incubation, any zone of inhibition around the disk is considered as susceptible
12. CAMP test • Principle: • Group B streptococci produce extracellular protein (CAMP factor) • CAMP act synergistically with staph. -lysin to cause lysis of RBCs • Procedure: • Single streak of Streptococcus to be tested and a Staph. aureus are made perpendicular to each other • 3-5 mm distance was left between two streaks • After incubation, a positive result appear as an arrowhead shaped zone of complete hemolysis • S. agalactiae is CAMP test positive while non gp B streptococci are negative
13. CAMP test
14. Differentiation between -hemolytic streptococci • The following definitive tests used to differentiate between S. pneumoniae & viridans streptococci • Optochin Test • Bile Solubility Test • Inulin Fermentation
15. Optochin Susceptibility Test • Principle: • Optochin (OP) test is presumptive test that is used to identify S. pneumoniae • S. pneumoniae is inhibited by Optochin reagent (<5 µg/ml) giving a inhibition zone ≥14 mm in diameter. • Procedure: • BAP inoculated with organism to be tested • OP disk is placed on the center of inoculated BAP • After incubation at 37oC for 18 hrs, accurately measure the diameter of the inhibition zone by the ruler • ≥14 mm zone of inhibition around the disk is considered as positive and ≤13 mm is considered negative • S. pneumoniae is positive (S) while S. viridans is negative (R)
16. Optochin Susceptibility Test Optochin resistant S. viridans Optochin susceptible S. pneumoniae
17. Bile Solubility test • Principle: • S. pneumoniae produce a self-lysing enzyme to inhibit the growth • The presence of bile salt accelerate this process • Procedure: • Add ten parts (10 ml) of the broth culture of the organism to be tested to one part (1 ml) of 2% Na deoxycholate (bile) into the test tube • Negative control is made by adding saline instead of bile to the culture • Incubate at 37oC for 15 min • Record the result after 15 min
18. Bile Solubility test • Results: • Positive test appears as clearing (trong) in the presence of bile while negative test appears as turbid (đục) • S. pneumoniae soluble (hòa tan) in bile whereas S. viridans insoluble
19. Differentiation between -hemolytic streptococci Hemolysis Bacitracin CAMP test sensitivity S. pyogenes  Susceptible Negative S. agalactiae  Resistant Positive Differentiation between -hemolytic streptococci Hemolysis Optochin Bile Inulin sensitivity solubility Fermentation S. pneumoniae Sensitive Soluble Not ferment (≥ 14 mm) Viridans strep Resistant Insoluble Ferment (≤13 mm)
20. Outline of differentiation between Gram-Positive cocci e.g. S. epidermidis