A reverse transcriptase-loop mediated isothermal amplification assay (RT-LAMP) for rapid detection of bovine viral diarrhea virus 1 and 2

Bovine viral diarrhea virus (BVDV) is a pathogen that infects cattle, and is globally important. It causes substantial financial losses to the livestock industry. In the current study, a one-step reverse transcriptase-loop-mediated isothermal amplification (RT-LAMP) assay was set up for rapid and efficient detection of BVDV. For this purpose, four primers were designed to recognize six distinct regions on the target RNA based on a highly conserved sequence in the 5΄ UTR of the BVDV genome. Eighty blood specimens were collected from bovines suspected to suffer from BVDV infection, and were tested in parallel by RT-LAMP and RT-PCR. Twenty four of these samples were positive by RT-LAMP, while twenty were positive by RT-PCR. The RT-LAMP detection limit was estimated to be approximately 70PFU /mL of virus. Comparison of RT-PCR with RT-LAMP in this study revealed the recent developed RT-LAMP a highly sensitive and specific for BVD virus detection in the clinical samples.