一步法RT-PCR检测猪繁殖与呼吸综合征病毒的方法建立及应用

Establishment and Application of One-step RT-PCR Detected Porcine Reproductive and Respiratory Syndrome Virus

根据GeneBank公布的猪繁殖与呼吸障碍综合征病毒(PRRSV)美洲型毒株VR-2332序列设计了一对特异性引物P1/P2,扩增片段为508bp,从而建立了检测PRRSV的一步法RT-PCR蛤测方法,特异性试验表明,该引物均不能扩增其它常见的与繁殖障碍有关的病毒基因。敏感性实验表明,该引物可以检测到10-5TC ID50的病毒含量。利用该方法对青海某猪场中20份临床上疑似PRRS的组织进行检测,其中16份样品呈PRRSV阳性结果,阳性率为80%,表明建立的该方法完全可以快速检测组织中的PRRSV。

According to the sequence of American strain VR-2332 of porcine reproductive and respiratory syndrome virus（PRRSV） published by the GenBank , a pair of specificity primes P1/P2 were designed and the length of ampIify fragment was 508 bp to establish the one-step RT-PCR detected PRRSV. The results of specificity test showed that the other common virus gene which related reproductive disorder showld be not amplified by this pair of primes. The results of sensitive test showed that the 10^-5 TCID50 of virus should be detected by this pair of primes, 20 Samples of tissue of pig which was clinically suspected of PRRS in a pig farm of Qinghai province were detected by means of this method, and 16 samples were presented positive result for PRRSV and the posicive rate was 80%, suggesting that the PRRSV in tissue of illness pig should be fast detected by means of the established method.