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

TEST PROPER

• Deliver 25 µl of antisera to the row of wells for each virus.
• Add 25 µl if antigen to each well having antisera.
• Add 50 µl of complement containing 3 units. Shake the plate well and incubate at 37 ⁰ C for 1 h.
• In the mean time prepare indictor system (4 MHD 50 per ml of haemolysin and 1.4% final concentration of standardized RBC) incubate at 37 ⁰ C for minutes
• After incubation is over for both plates as well as sensitized cells, 25 µl of indicator system is added to all the wells.
• The plate is shaken gently, covered, sealed and incubated at 37⁰C for 1 h and then kept at 4 ⁰ C overnight.
• Controls
  • Sensitised cell control - 0.1 ml VBD and 25 µl l indicator system
  • Complement control - 50 µl of VBD, 50 µl of indicator system
  • Serum control - 25 µl of VBD, 25 µl complement and 25 µl indicator system.
  • Antigen control - 25 µl VBD, 25 µl antigen, 50 µl complement and 25 µl indicator system.