Rapid visual detection of bovine viral diarrhea virus (BVDV) using recombinase polymerase amplification with SYBR green I.

Abstract

Background: Bovine diarrhea virus (BVDV) is considered to be the most common pathogen causing severe diarrhea in cattle worldwide and can cause Bovine viral diarrhea (BVD). Clinical manifestations of fever, diarrhea, ulcers, and abortions, resulting in significant economic losses to the cattle industry. The development of an efficient, rapid and sensitive assay suitable for field conditions is of great significance for its early detection. Recombinase polymerase amplification (RPA) is a novel nucleic acid amplification method that has been widely used in the diagnosis of infectious diseases.

Results: We developed a rapid assay (RPAS) combining RPA with SYBR Green I for the detection of BVDV. The BVDV RPAS assay was performed at 37 °C in 25 min. The minimum detection limit of the RPAS assay is 1 × 10⁹ copies/µL in sunlight and 1 × 10⁵ copies/µL in ultraviolet light, and there is no cross-reactivity with other viruses that cause gastrointestinal and respiratory infections in cattle. The coincidence rate of BVDV RPAS in clinical samples was higher than that of PCR.

Conclusions: The BVDV RPAS assay established in this study has high sensitivity and specificity, and is expected to be a powerful tool for the prevention and control of BVD.