Abstract: Senecavirus A(SVA) is a nonenveloped, single-stranded RNA virus, which can cause vesicular lesions in sows and newborn piglet death. In order to establish a rapid assay for detection of SVA, we compared the genetic sequences of 40 SVA genes published on NCBI, and the best primers was selected from three pairs of primers which designed based on the conserved regions of sequences. The RT-PCR detection method of SVA was established successfully after the optimization of primer concentration, annealing temperature, extension time and cycle number. SVA, Encephalomyocarditis virus (EMCV), foot-and-mouth disease virus (FMDV), pseudorabies virus (PRV), porcine reproductive and respiratory syndrome virus (PRRSV), swine fever virus (CSFV), porcine epidemic diarrhea virus (PEDV) were detected by RT-PCR, which showed a good specificity with no cross-reaction. The sensitivity test was carried out as well, and sensitivity of the detection could reach up to 1 TCID₅₀, which shows it is more sensitive than the methods reported in foreign literatures. One hundred samples of pig tissue were detected using this method, and the positive rate was 2%. The establishment of this method, with its great specificity, high sensitivity and ideal reliability, could provide an effective technical means for SVA detection and epidemiological investigation.