A microtiter test for detecting and titrating noncytopathogenic bovine viral diarrhea virus

Bovine cells free of noncytopathogenic bovine viral diarrhea virus (NC-BVDV) treated with polyriboinosinic acid : polyribocytidylic acid (poly I:C) were protected against challenge with vesicular stomatitis virus (VSV), whereas NC-BVDV-infected cells treated with poly I:C were not protected against VSV. An assay based on the ability of NC-BVDV to inhibit poly I:C protection of cells against VSV was developed and is herein referred to as PINBA (poly I:C for NC-BVDV assay). Noncytopathogenic BVDV was titrated as cytopathogenic strains except that several days after infection with NC-BVDV, the cultures were treated with poly I:C and VSV. Titration endpoints were reached 24 hours later. PINBA was standardized for amount of VSV, time of addition of poly I:C, and time NC-BVDV had to be present to obtain stable titration endpoints. PINBA also was useful for titrating virus neutralizing antibodies. Compared with the fluorescent antibody test, PINBA was less subjective for detection of NC-BVDV. Compared with the interference test in which NC-BVDV infected cultures are challenged with a cytopathogenic strain of BVDV, PINBA was more reliable. The technique described herein is a simple and practical microtiter method for titrating NC-BVDV and virus neutralizing antibodies and for the presumptive detection of NC-BVDV.