Contamination of Cell Cultures with Bovine Viral Diarrhea Virus (BVDV)

Abstract

The incidence of contamination of cell strains used in biological and virological studies and of fetal calf sera (FCS) manufactured by Russian and foreign companies used for cell culturing with noncytocidal bovine viral diarrhea virus (BVDV; Pestivirus, Flaviviridae) was analyzed. The virus was detected by reverse transcription PCR and indirect immunofluorescence with monoclonal antibodies to BVDV virion envelope glycoprotein in 25% of 117 cell strains and 45% of 35 tested FCS lots. The virus multiplied and persisted in a wide spectrum of human cell strains and in monkey, swine, sheep, rabbit, dog, cat, and other animal cells. The levels of BVDV genome RNA in contaminated cell cultures reached 10(2)-10(3) g-eq/cell and in serum samples 10(3)-10(7) g-eq/ml. These facts necessitate testing of cells and FCS for BVDV reproduced in cells without signs of infection detectable by light microscopy. The molecular mechanisms of long-term virus persistence in cells without manifestation of cell destruction are unknown.

Application of next generation sequencing technology on contamination monitoring in microbiology laboratory

“…The sequences of bovine diarrhea virus were only found on the biosafety cabinet in Room 1, which was used for viral isolation and culture experiments ( Figure 2). The existence of bovine diarrhea viral nucleic acids in fetal bovine serum (FBS) was further confirmed by PCR methods (data not shown) [18], which may represent contamination of bovine serum used for cell culture as reported also in previous studies [18][19][20][21].…”

“…(2) Experimental materials. For example, the fetal bovine serum was confirmed to be contaminated by the bovine viral diarrhea virus [18][19][20][21]. Besides that, the reverse transcriptase which derived from murine leukemia virus [27][28][29], and the recombinant protein expression system which involves the use of viral vectors are potential sources of contamination [30][31][32].…”

Genetic Diversity of Brazilian Bovine Pestiviruses Detected Between 1995 and 2014

“…The source of the contamination is not known, although it probably arose from contaminated FBS. Contaminations of FBS and cell cultures by BVDV‐1, BVDV‐2 and HoBi‐like pestivirus have been reported (Bolin and Ridpath, ; Xia et al., ; Uryvaev et al., ; Oliveira et al., ; Gagnieur et al., ; Zhang et al., ). In the present study, twelve isolates from four different genogroups (BVDV‐1a, 1d, 2c and HoBi‐like pestivirus) were detected in either cell culture or FBS.…”

2.5 Quality Control Essentials in Human Cell Culture: Cell Line Cross-contamination and Microbiological Infections

“…This usually leads to the contamination with airborne bacteria, yeasts, or fungi that rapidly overgrows the eukaryotic cells and can be detected shortly after introduction. On the other hand, a microbial contamination can already be present in the primary cells originating from infected tissue or in cell culture media and supplements due to contaminated raw material [20]. Another source of infection can be the transmission of bacteria or viruses from one cell culture to another cell culture (cross-contamination).…”