Abstract

CORDERO CAMACHO, Claudia P et al. Detection of seven viruses and Mycoplasma in fetal bovine serum by real time PCR. Rev Colom Cienc Pecua [online]. 2011, vol.24, n.4, pp.585-597. ISSN 0120-0690.

Objective: real  time  PCR  analysis for the  detection  of seven bovine  pathogenic viruses: Bovine Adenovirus  (BAdV), Bovine  Viral Diarrhea  Virus  Types  1  and  2  (BVDV-1  and  BVDV-2),  Bovine Respiratory Syncytial Virus  (BRSV),  Vesicular Stomatitis Virus (VSV), Bovine  Parainfluenza  Virus  3 (BPIV-3), Bovine Herpes Virus-1 (BoHV-1), and Mycoplasma was conducted using fetal bovine serum (FBS, MICROGEN^®) obtained in Colombia,  aiming  to  include it as part of the serum  quality control. Methods: bovine derived MDBK and human derived HEp-2 cell lines were cultured with the test serum for 21 days, collecting supernatant and cellular samples every 7-days. Once DNA and RNA were extracted, the later was converted into cDNA and both samples were subjected to real time PCR using specific primers and Resolight^® (DNA-binding fluorescent dye). Standard curves were generated using serial dilutions of cloned  specific  viral sequences. Accurate amplification  and  high  efficiency was demonstrated  in these reactions. Results: realtime PCR amplification did not show a persistent increase of viral counts in cultures during the 21-day follow-up. However, for vesicular stomatitis virus, a transient increase was observed at 7 and 14 days in both cell lines, but considered as not conclusive for viral presence. Conclusions: real time PCR analysis showed to be a suitable method for viral detection in fetal bovine serum samples and through this method no consistent viral or mycoplasma presence was detected in the MICROGEN^® fetal bovine serum.

Keywords : bovine virus; fetal bovine serum; real time PCR; viral detection.

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