Development of a CRISPR/Cas12a-based fluorescent detection system of Senecavirus A

Abstract Background Senecavirus A (SVA) was found in 2002 and could cause porcine idiopathic vesicular disease (PIVD) which symptoms were similar to vesicular diseases resulting in increased difficulty of a field diagnosis. However, traditional molecular diagnosis failed to reconcile cost, instrument, sensitivity, and efficiency. Methods In this study, we integrated pre-amplification and three kinds of sensor systems with CRISPR and therefore established an SVA diagnosis platform with highly adaptable and ultra-sensitive advantages. This diagnosis which the whole process should not exceed four hours consisted of three steps: nucleic acid extraction, pre-amplification, and fluorescent signal capture. Results This method showed no cross-reaction with other 10 swine viruses; its limit of detection was as low as one copy/reaction of SVA; its accuracy for clinical sample diagnosis was 100%. Conclusions In addition, this diagnosis used common instruments which would be beneficial to remote testing sites or poorly equipped laboratories to promote a large-scale screening of this epidemic. Overall, this diagnosis enlarged diagnostic tools of SVA and had the potential to play a positive role in the control of PIVD.