摘要
根据GenBank上发表的猪流行性腹泻(PEDV)和猪传染性胃肠炎(TGEV)基因序列,针对其PEDVM(膜蛋白)基因保守区及TGEV的S基因(纤突蛋白基因)5′端保守区,各设计一对引物,可特异扩增出目的条带大小分别为467bp和1062bp。用上述两对引物对同一样品中的PEDV和TGEV可进行鉴别检测,对猪的其它病毒和细菌的PCR扩增结果均为阴性。敏感性测定结果表明:该双重PCR方法能检出PEDV1pg、TGEV0.1pg的模板。同时采用建立的多重RT-PCR及韩国引进的PEDV和TGEV病毒抗原快速诊断试剂盒检测结果显示:此多重RT-PCR方法特异性强,且较快速试剂盒更加敏感。应用多重RT-PCR对158份临床猪粪样的检测结果表明:我国很多猪场普遍存在TGEV和PEDV,尤其以PEDV污染更为严重,感染率达53.2%,但双重感染率较低,仅为4.4%。
A multiple reverse-transcriptase-PCR （RT-PCR） procedure was developed for the simultaneous detection of PEDV（PEDV） and TGEV（TGEV） in pigs .The membrane gene of PEDV and spike gene of TGEV were chosen as targets. The PCR products of PEDV and TGEV had molecular sizes of 467 bp and 1 062 bp, respectively. Primers from PEDV did not react with TGEV and other porcine viruses. The multiplex RT-PCR was able to detect 1 pg tissue culture-infective doses of PEDV or 0.1 pg TGEV with each of the primer sets for PEDV and TGEV, respectively. The comparison results of clinical samples using multiplex RT-PCR and Antigen Rapid Test Kit revealed： our method is differential and sensitive. The RNA of PEDV and TGEV were detected directly in intestinal and faecal samples from pigs infected experimentally with either virus. The detect results of 158 shares of clinical samples showed： PEDV and TGEV are ubiquitous in China and PEDV is severe （53.2 % ）. Duple infection of PEDV and TGEV is only 4.4 %.
出处
《中国预防兽医学报》
CAS
CSCD
北大核心
2006年第4期431-435,共5页
Chinese Journal of Preventive Veterinary Medicine
关键词
猪流行性腹泻病毒
猪传染性胃肠炎病毒
猪呼吸道冠状病毒
双重RT-PCR
porcine epidemic diarrhea virus （PEDV）
transmissible gastroenteritis virus （TGEV） porcine respiratory coronarivus
multiplex RT-PCR
