| According to the gene sequences in GenBank of PRRSV ORF7 N gene and JEV NS1 gene, two pairs of specific primers were designed for amplifying the two specific fragments of PRRSV and JEV respectively. After optimization of annealing temperature and primers concentrations, dual one-step RT-PCR detection kit for PRRSV and JEV was development. Two specific bands of 245 bp (PRRSV), 504 bp (JEV) were amplified. The sensitivity and specificity tests showed that the dual one-step RT-PCR was highly sensitive in PRRSV 0.2 ng/L, JEV 0.4 ng/L. No band was amplified from CSFV, PRV, PCV-2, PPV by the dual one-step RT-PCR. 52 clinical samples were detected by the dual one-step RT-PCR detection kit for PRRSV and JEV. The results revealed that the dual one-step RTPCR detection kit was sensitive, specific and reproducible and it could be used to detect rapidly PRRSV and JEV. |
