“…Control wells were coated with PBS. After washing the plate 5 times with PBS-0.05% plification were conducted (20 seconds at 94 ЊC for denaturing, 30 seconds at 55 ЊC for annealing, and 30 Tween 20 (PBS-T), 50 mL of test serum diluted 1:10 in PBS-0.5% nonfat milk was added to each of 2 antigen seconds at 72 ЊC for extension, with a final extension of 5 minutes) using a Perkin-Elmer 9600 thermocycler and control wells. The plate was incubated for 2 hours at 37 ЊC, then washed 5 times with PBS-T. VLP-reactive (Perkin-Elmer, Oak Brook, IL).…”