A reverse transcription loop-mediated isothermal amplification method for rapid detection of bovine viral diarrhea virus

A reverse transcription loop-mediated isothermal amplification (RT-LAMP) assay was developed and optimized to detect bovine viral diarrhea viral (BVDV) RNA. The RT-LAMP assay is highly sensitive and able to detect 4.67 × 100 copies of BVDV RNA. Additionally, the RT-LAMP method is capable of detecting both genotypes of BVDV. No cross-reaction with other bovine viruses was observed. The ability of RT-LAMP to detect BVDV RNA from bovine fecal swabs was also evaluated. Of the 88 fecal swabs, 38 were found to be positive by RT-LAMP assay, whereas 39 were positive by real-time RT-PCR. Taken together, the BVDV specific RT-LAMP method is highly specific and sensitive and can be used as a rapid and direct diagnostic assay for testing clinical samples.

► A set of primers specific to the BVDV genome was designed and used in the RT-LAMP. ► Both genotypes of BVDV can be detected by the RT-LAMP method. ► The RT-LAMP is a rapid, sensitive and specific diagnostic assay.