Studies on Detection of Porcine reproductive and respiratory syndrome virus in Tissue Sample by Reverse Transcription Polymerase Chain Reaction
- 生命科学－已发表论文 
根据猪繁殖和呼吸综合征病毒(PRRSV)已发表的核苷酸序列 ,在其核衣壳蛋白基因 (ORF7)保守区设计了一对跨幅约 380bp的特异性引物N1/N2 ,对已知PRRSVJK10 0 1毒株和PRRS临床病料进行RT PCR扩增 ,均扩增出约 380bp的特异片段。用此方法检测 5份疑似PRRS病例肺组织病料 ,结果均为阳性 ,对 2个阳性样品扩增产物进行克隆、测序 ,全长均为 372bp ,与PRRSVVR 2 332毒株ORF7同源性为 95%,证实为PRRSV的核衣壳蛋白基因。结果表明 ,建立的RT PCR检测PRRS临床组织病料特异、快速 ,可用于PRRS临床病料检测According to the published sequence of PRRSV genome, a pair of specific primers (N1/N2) was designed based on the gene encoding the N protein of PRRSV. The region between the primers was about 380 bp. A reverse transcription-polymerase chain reaction (RT-PCR) was developed for the detection of PRRSV in clinical lung tissue samples. Five clinical lung samples were detected by RT-PCR and all were identified as PRRSV positive, two of them were further cloned and sequenced. The result showed that both genes are 372 bp in length and shared very high homology to the ORF7 gene of PRRSV VR-2332 strain.In conclusion, the RT-PCR is specific and rapid and can be applied to diagnose PRRS clinically.