A RT-PCR Assay for Quantitative Detection of Bovine Viral Diarrhea Virus

  Objective   A rapid detection and quantification method for bovine viral diarrhea virus (BVDV) was established.

  Method   For the methodology development, specific primers and probes were designed based on the target regions of the BVDV 5′UTR gene published by GenBank. The RNA of in vitro transcription viruses was used as the absolute quantitative standard. Reaction conditions of the fluorescent quantitative RT-PCR were optimized.

  Result  The newly developed assay had a minimum detection limit of 5.0267copies/μL and an intragroup variation coefficient of less than 1% with high repeatability and specificity. Other than BVDV, it amplified no viral nucleic acids of viruses such as swine fever and foot-and-mouth diseases.

  Conclusion  The established fluorescence quantitative RT-PCR method was highly sensitive, specific, and repeatable in detecting and quantifying BVDV. It appeared appropriate for early diagnosis of bovine viral diarrhea.