ANTI-STREPTOLYSIN O (ASL)

INTRODUCTION Streptolycin O is a haemolytic toxin produced by most pathogenic streptococcus strains. The toxin immunogenic, and serum levls of ASL are often raised following recent streptococcal infection. Measurement of ASL serum titres can therefore be useful in the diagnosis of streptococcal infection, particularly in cases of rheumatic fever and glomerulonephritis. Streptolysin O antibodies may persist for variable periods post infection and so can be detected in healthy subjects.

MATERIAL PROVIDED 1. Latex reagent 2. Pre-diluted positive serum control 3. Pre-diluted negative control serum 4. Glycine saline buffer (pH 8,2)

SAMPLE COLLECTION AND STORAGE Collect 5-10 mL venous blood in to sterile glassware without anticoagulant. Alloe to clot for several hours at room temperature. Formation of the clot must be complete. Separate serum and store at 2-8oC. It is preferable to test these samples within 2-3 days of storage.

SCREENING TEST 1. Bring all reagent and test specimens to ambient temperature. 2. Prepare a 1:20 primary dilution of test serum by adding 50L of serum to 1mL of glycine saline solution provided. 3. Place 50L of diluted serum on to the test slide. 4. Add 1 drop of gently mixed latex reagent. 5. Mix the tow drop together with a clean stirrer and spread out to the edge of the test area. 6. Rock the slide gently and observe for macroscopic agglutination under a direct light source. 7. Read at 3 minutes.
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SEMIQUANTITATIVE TEST
the serum to be titrated can be serially diluted (from the primary dilution) in the glycine saline solution as follow :

Secondary dilution

Primary Dilution (mL) 0,2 0,1 0,1 0,1 0,1

GBS (mL) 0,1 0,1 0,2 0,3 0,4

Final ASL Titre Dilution (IU/mL) 1:30 1:40 1:60 1:80 1:100 300 400 600 800 1000
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Each dilution is then tested as described in point (3) to (4) above.

INTERPRETAION Negative reaction : the latex reagent maintains its original homogenenous milky appearance. Positive reaction : agglutination of the latex particles be visible, varying between a fine granulation againts a milky background and a gross clumping againts a clear background.

SEMIQUANTITATIVES METHOD The ASL titre should recorded as the greatest dilution showing a clear positive agglutination. A conversion of the end point in to international unit may be made by multiplying by the treshold sensitivity of the latex (10mL) EXAMPLE Last positive dilution 1:60 ASL titre = 60 x 10 IU/mL = 600 IU/mL
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