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hemolytic
divided into serogroups A through O contains an important surface component called FIMBRIAE
arise near the plasma membrane and project through the cell wall and capsule contains LIPOTECHOIC ACID important for adherence to human epithelium for initiation of infection
M Antigens
major virulence factor
strongly anti-phagocytic
R Antigens
no known biologic role
Scarlet Fever
pharyngeal infection caused by erythrogenic toxin by GAS produces characteristic rash develops on the 2nd day of illness results in hyperkeratosis with subsequent peeling
Skin Infection
Impetigo lesion that itches then crusts over and heals Cellulitis subcutaneous infection - characterized by warm, red, tender area that is slightly swollen
Skin Infection
Erysipelas distinct cellulitis syndrome - involves the face - associated with pharyngitis - characterized by toxicity and high fever - fatal if untreated
Glomerulonephritis
may occur after pharyngitis or skin infection inflammation of the glomeruli - small blood vessels in the kidney Primary causes are ones which are intrinsic to the kidney Secondary causes are associated with certain infections
Streptolysin O (SLO)
bacterial strains produced by
Streptococcus pyogenes
released during infection which is an indication of the production of antibody in reduced rate causes the lysis of the red and white blood cells
Streptolysin O (SLO)
oxygen labile hemolytically inactive in oxidized form
Streptolysin O (SLO)
low concentrations of cholesterol and related sterols is inhibited by their biological activity
Streptolysin O (SLO)
Cardiotoxic may cause interstitial myocarditis in experimental animal - causes systolic arrest of perfused mamalian heart - caused by inducing the release from the atria of ACETYLCHOLINE, that poisons the ventricles
Streptolysin O (SLO)
f site - 2 cystine residues - responsible for the attachment of the molecule to red blood cell t site - hemolytic event
Streptolysin O (SLO)
membrane cholesterol binding site exogenous cholesterol inhibits toxic action
Anti-Streptolysin O
-valuable and reliable indicator of Streptococcal infection
Streptolysin O
- antigenic - elicits formation of antibody that effectively neutralize hemolytic reaction
Streptolysin O
Oxygen labile Antigenic Synthesized by growing Streptococcus
Streptolysin S
Oxygen stable Non antigenic peptide Sythesized both by growing and resting cells
Causes necrosis of the liver and kidney tubules and massive intracellular hemolysis upon injection intravenously
Induce chronic arthritis upon injection intra- articularly
Neutralization test
detect ASO in serum ASO can be specifically fixed to SLO, in vitro hemolytic activity is neutralized by doubling dilution, estimates the amount of antibody with the presence of a constant dose of SLO, completely inhibits hemolysis
Variations: age
severity of infection previous exposure to Streptococcal infection individuals ability to respond immunologically to the toxin
ASO Titers
in children's, fluctuates from 5 125 Todd units 30% rise over the previous level is considered significant. increases due to rheumatic fever and glomerulonephritis is seen during symptomfree period preceding attack of the illness
ASO Titers
in rheumatic fever 300 to 2, 500 Todd units - maintained at high levels for 6 months increased amount is found in: -Scarlet fever - Cholera minor - Tuberculosis disease - Pneumococcal pneumonia - gonorrhea
that amount of toxin will completely hemolyze 0.5 ml of a 5% suspension of rabbit blood cells, measured in Todd units
MATERIALS
Saline 0.85% Streptolysin O buffer: 7.4 gm sodium chloride 3.17 gm potassium 1, 081 gm sodium phosphate add 1, 000 ml of distilled water
STREPTOLYSIN O
if in dehydrated form must be rehydrated before use once rehydrated the solution should not be subjected to vigorous shaking, and must be used within a period of 1 hour or discarded active reagent is subject to inactivation by oxidation
5% suspension of fresh human RBC (grp O) Rabbit BBC equally sensitive to SLO cells must be washed 3 times in diluent buffy coat must be removed Final centrifugation - 1, 500 rpm for 10 minutes Packed RBC = 5% suspension Final suspension in SLO buffer
TEST TUBE
must be 12 x 100 mm round bottom
PROCEDURE:
1. Prepare dilutions of fresh or inactivated serum, using SLO buffer as diluent:
PROCEDURE:
1. Prepare dilutions of fresh or inactivated serum, using SLO buffer as diluent:
1:10 - 0.5 ml of serum + 45 ml of buffer
The first 2 serum dilutions are usually sufficient for preliminary titrations
PROCEDURE:
2. Set up the test according to the protocol given TABLE 9.1 PAGE 188
INTERPRETATION:
the ASO titer expressed in Todd units is the reciprocal of the serum dilution that completely neutralizes the SLO.
before reporting results, always ensure that the controls give the expected results
PRINCIPLE:if polystyrene latex particles are coated with SLO antigen, visible agglutination will be exhibited in the presence of the corresponding ASO antibody
0.9% NaCl solution -contains sodium azide as preservative Positive (+) control serum - contains at least 200 U/ml of ASO - should exhibit visible agglutination at the end of 3 min test period
MATERIALS
Negative (-) control serum -Prediluted serum with less than 100 U/ml of ASO
Glass slides with 6 wells
ADDITIONAL MATERIALS
1. 2. 3. 4. 5. 6. 7. Applicator sticks Timer 12 x 75 mm test tubes Pasteur pipettes and rubber bulb Serologic pipettes and safety bulb 50 l disposable pipettes and safety bulb High-intensity direct light
Add 2 drop of patient serum Cover the tube and mix the dilution thoroughly by inverting the tube several times Label 1 division of the 6-cell slide for the positive control, negative control, and the respective patient sera Pipette 50 l of the controls and patient sera
Mix each specimen with a separate applicator stick. Spread the mixture evenly on the cell. Rotate the slide for exactly 3 minutes. Examine immediately with a bright source of direct light
INTERPRETATION
DISCUSSION
False Positive reactions - result of bacterial contamination - if the reaction is absorbed after 3 minutes - Lipemic serum or plasma
DISCUSSION
RHEUMATIC FEVER or GLOMERULONEPHRITIS - titer with 200 U/ml or greater
elevated titer should be retested over a period of 4 to 6 weeks to plot the course of titer