A Method to Detect Bovine Viral Diarrhea Virus Contamination in Cell Cultures using Immunoperoxidase Staining

Abstract

Certain tests in veterinary diagnostic laboratories are dependent on the use of cell cultures free of contamination with noncytopathic (nc) bovine viral diarrhea virus (BVDV). Such tests include virus and chlamydia isolations, serum neutralization assays, and the microplate virus isolation technique for detection of BVDV in persistently infected animals. 1,3 Cells infected with ncBVDV appear morphologically

Single-Tube Single-Enzyme Reverse Transcriptase PCR Assay for Detection of Bovine Viral Diarrhea Virus in Pooled Bovine Serum

“…Virus stocks were propagated in MDBK cells. The microplate virus isolation assay was performed as described previously (1,5).…”

Failure to Detect Bovine Viral Diarrhea Virus in Necropsied Farm-Raised White-Tailed Deer (Odocoileus Virginianus) in Pennsylvania

“…The supernatant was filtered using 0.2 μm membrane filter and inoculated onto 25 cm 2 tissue culture flasks and 4-chambered tissue culture slides of Madin-Darby bovine kidney cells (MDBK) cells. Maintenance medium used was 2% fetal bovine serum (BVDV-free based on virus culture, immunoperoxidase staining of cell monolayers, and realtime RT-PCR) in minimal essential medium (MEM) with antibiotics (kanamycin, gentamicin, penicillin, streptomycin, amphoterocin) 4 . Positive cell-culture controls consisted of MDBK cells inoculated with noncytopathic BVDV-NY-1.…”

“…Maintenance medium used was 2% fetal bovine serum (BVDV-free based on virus culture, immunoperoxidase staining of cell monolayers, and realtime RT-PCR) in minimal essential medium (MEM) with antibiotics (kanamycin, gentamicin, penicillin, streptomycin, amphoterocin). 4 Positive cell-culture controls consisted of MDBK cells inoculated with noncytopathic BVDV-NY-1. Cell culture flasks were incubated in 5% CO 2 at 37uC and observed for cytopathic effects for 6 days.…”