Systematic Veterinary Virology

PROCEDURE

• Take three test tubes and mark them as A, B and C.
• Dilute the positive serum to 1:10, 1:100, and 1:1000 by adding diluent buffer
• Take two clean glass slides and in one slide make two separate circles, one for the positive test serum and the other for the negative test serum and mark them ‘+ve’ and ‘-ve’. In second slide make three circles for 1:10, 1:100, and 1:1000 dilution tests and label them accordingly
• Transfer 20 μl of sensitized latex to each circle of both the slides.
• Add 20 μl of positive test serum into the circle marked as ‘+ve’ and add 20 μl negative test serum into the circle marked as ‘-ve’. Similarly add 20 μl each of the diluted sera into the respective circles.
• Thoroughly mix the sensitized latex and serum suspension by using separate toothpicks or tips.
• Rock the slide gently for up to 2 minutes and avoid mixing the contents of the different circles.
• Agglutination occurs in the positive test circle, whereas milky suspension remains in the negative test circle.