VMC 221: Veterinary Immunology and Serology (1+1)

MATERIALS

• Normal saline
  • 0.85 g sodium chloride in 100 ml of distilled water.
• Alsever’s solution – anticoagulant used during collection of blood for preparation of erythrocyte suspension.
  • Sodium chloride - 4.2 g
  • Sodium citrate - 8.0 g
  • Dextrose - 20.5 g
  • Citric acid - 0.55 g
  • Distilled Water - 1000 ml
  • Sterilize at 10 lbs for 10 minutes and store at 4 ⁰ C
• 1% chicken erythrocyte suspension
  • Collect blood from four numbers of 6-week-old chickens in Alsever’s solution in the proportion of 1.1 ml of Alsever’s solution for each ml of blood collected. Wash the erythrocytes thrice by gentle centrifugation in sterile normal saline at 1500 rpm for 5 minutes Prepare 1% suspension by adding 99 ml of sterile normal saline to 1 ml of the packed erythrocytes and store at 4 ⁰ C
• Microtitre plates – ‘V’ bottom.
• Micropipette – 20-200 μl  and tips.
• Antigen: Allantoic fluid collected from embryonated eggs infected with Newcastle disease virus.
• 4 HA units
  • 4 HA = HA titre / 4
  • Example: When HA titre = 128, 4 HA = 128 /4 = 32