Novel M-PCR assay for simultaneous detection of gastroenteritis associated viruses

Abstract

Abstract Bovine Viral diarrheal virus (BVDV), bovine corona virus (BCV), and bovine picobirnavirus (BPBV) group I and II are most common viral infections in cattle that cause a huge economic loss across the world. A rapid and novel multiplex-PCR assay (M-PCR) was developed for simultaneous detection of BVDV, BCV and BPBV. Three primer pairs specific for each virus were used in the assay. Out of these three primer pairs, two pairs from M and 5’ UTR gene specific primer for BCV and BVDV respectively were self designed and .used for amplification. To evaluate the reliability of the multiplex assay, 300 buffalo faecal samples were tested by multiplex PCR assay. The results showed that 54(18%) samples were detected positive with multiple viruses including 50 samples were infected with BVDV, twenty eight samples were infected with BCoV and thirty nine samples were found infected with BPBV by M-PCR assay. In conclusion the developed assay is found highly specific, sensitive, high throughput and quick method for the detection of three viruses in a single reaction tube. This multiplex assay is successful tool that can be used for quick detection of these three viral infections for epidemiological investigation and in screening of clinical samples. To the best of our knowledge, this is the first time reported multiplex assay for the detection of BVDV, BCoV and BPBV throughout the world. This novel assay will be helpful for detecting mixed infections of GITs.