猪传染性胃肠炎病毒和猪流行性腹泻病毒二联RTPCR检测方法的建立.pdfVIP

中国农学通报 2010,26(23):26-29 Chinese Agricultural Science Bulletin 猪传染性胃肠炎病毒和猪流行性腹泻病毒 二联RT-PCR检测方法的建立 李敬双，于 洋 （辽宁医学院，辽宁锦州121001） 摘 要：参照国内外已发表的猪传染性胃肠炎病毒（TGEV ）和猪流行性腹泻病毒（PEDV ）基因序列及其 相关的RT-PCR 检测方法，根据TGEV S 蛋白基因和PEDV S 基因各设计一套特异性通用引物，扩增目的 带分别为426 bp 和584 bp 。通过对相关病毒检测，建立了TGEV 和PEDV 通用型二联RT-PCR 检测方 法。该方法具有快速、敏感、特异等优点，可为TGEV 和PEDV 的检测、流行病学调查及疫苗使用等奠定 基础。 关键词：猪传染性胃肠炎病毒；猪流行性腹泻病毒；反转录聚合酶链式反应 中图分类号：S859 文献标志码：A 论文编号：2010-1396 EstablishmentofTGEVandPEDVbyMultiplexRT-PCR Li Jingshuang, Yu Yang （LiaoningMedicalUniversity, Jinzhou Liaoning 121001） Abstract: According to the published gene sequence of transmissible gastroenteritis virus of pigs(TGEV) and porcine epidemic diarrhea virus(PEDV) and the detecting methods of RT-PCR reported, matrix gene of TGEV and nucleoprotein(NP) gene of PEDV were considered as the target gene sequences of multiple RT-PCR. A set of universe primers were designed for detecting TGEV and PEDV, respectively in the conseved S gene of TGEV and S gene of PEDV. The products of RT-PCR is 426 bp and 584 bp respectively. The optimal condition of multiple RT-PCR for detection of TGEV and PEDV by RT-PCR was determined through orthogonal assay. The specificity of the multipie primers were examined by RT-PCR using template extracted from other avian virus. The sensitivity of RT-PCR were also determined by a seral dilution of RNA from TGEV and PEDV. This study had paved the way for testing TGEV and PEDV, defining pathogenic potential, using vaccines against TGEV and PEDV and so on. Keywords:porcine transmissible gastroenteritis virus ；porcine epidemic diarrhea virus ；RT-PCR 0 引言