Establishment and application of dual TaqMan fluorescent quantitative PCR assay for Senecavirus A and encephalomyocarditis virus

ABSTRACT

In order to establish a method for rapid differential identification of Senecavirus A (SVA) and en-cephalomyocarditis virus (EMCV), two pairs of corresponding specific primers were designed based on the highly conserved 3D genes of SVA and EMCV. And two different fluorescent labeled TaqMan probes were used to establish a dual TaqMan real-time PCR method for simultaneous detection of these two viruses, and we also optimize the reaction conditions. The results showed that the minimum detection of the method was 760 copies/ micro L and 98 copies/ micro L for SVA and EMCV. respectively, and it can specifically detect SVA and EMCV, and there was no cross reaction with CSFV, PRRSV and PEDV. The established standard curves showed good linear relationship. Repeated experimental group and inter-group coefficient of variation were less than 5%. The results indicated that the dual-quantitative PCR established in this study has the advantages of convenience, rapidity, good specificity. high sensitivity and good repeatability .and can be used for simultaneous detection of SVA and EMCV.