Development of a real time loop-mediated isothermal amplification method for detection of Senecavirus A

Abstract

Senecavirus A (SVA), formerly known as Seneca Valley Virus (SVV), is one of causative agents of vesicular diseases in swine. Recently, the outbreaks associated with vesicular disease caused by SVA infection in pig herds have been reported in Brazil, USA, China, Thailand and Canada. Several molecular detection methods have been established to determine the infection of SVA, including real time reverse transcription PCR assay, nested PCR, a TaqMan-based qRT-PCR assay and RNA-based in situ hybridization method. In our study, an assay for the identification of SVA in pig herds using real time reverse transcription loop-mediated isothermal amplification (real time RT-LAMP) was developed. The limit of detection for the assay was 1 TCID₅₀/ml. One hundred and eighteen field samples from pigs were used to validate the assay for clinical application. Our result demonstrated that real time RT-LAMP assay is a cost-effective and highly specific and sensitive alternative for the rapid detection of SVA in clinical samples.

Keywords: Real time reverse transcription loop-mediated isothermal amplification; Senecavirus A; Vesicular disease.