摘要
根据猪瘟病毒5'非编码区(5'-UTR)设计特异性引物和Taqman探针,建立Taqman实时定量RT-PCR检测猪瘟病毒法。检测结果显示,该方法的灵敏度为1μl 10拷贝,在病毒拷贝数为1μl 108～101时,循环数(Ct)值与拷贝数对数呈现较好的线性关系,且重复性好,批间变异系数小于1%。用该方法检测猪繁殖与呼吸综合征病毒、牛病毒性腹泻病毒,结果均为阴性。用该方法检测采集自江苏和新疆的192份组织和血清样品,猪瘟病毒阳性率为71.9%;检测感染猪的不同脏器,发现在心、肺、肝、肾、脑、脾脏、淋巴结、腹水中均可以检测到猪瘟病毒,与常规RT-PCR方法相比,该方法敏感性更高。该方法的建立为猪瘟病毒的流行病学调查和定量提供了有效手段。
According to the conservative sequences located on the 5＇ untranslated region （5＇-UTR） of classical swine fever virus（CSFV） ,a pair of specific primers and Taqman probe were designed and synthesized respectively, and a Taqman real-time fluorescent quantitative reserve-transcribed polymerase chain reaction （real-time RT-PCR） for detecting the CSFV was established in this study. Test results showed that the method had a detection limit of 10 copies of target RNA per reaction, and there was a good linear relationship between Ct value and copy numbers in diluted samples. The variation between batches was less than 1%. The RNA of porcine reproductive and respiratory syndrome virus, bovine viral diarrhea virus were detected by the Taqman RT-PCR, and the results were all negative. The CSFV-positive rate was 71.9% in 192 samples collected from Jiangsu and Xinjiang areas. Real-time RT-PCR detection showed that the different organs of swine including hearts,lungs,livers, kidneys, brains, spleens, lymph nodes and ascites were CSFV-positive, indicating that the method were more sensitive and effective than traditional RT-PCR.
出处
《江苏农业学报》
CSCD
北大核心
2012年第4期783-786,共4页
Jiangsu Journal of Agricultural Sciences
基金
江苏省农业科技自主创新基金项目[CX(10)4129
CX(11)2143]
关键词
猪瘟病毒
实时定量RT-PCR
临床应用
classical swine fever virus （CSFV）
Taqman real-time RT-PCR
clinical application
