猪环曲病毒套式RT-PCR检测方法的建立及应用  

Establishment and application of a nest RT-PCR method for detection of porcine torovirus

作　　者：王赟[1] 周璐[1] 周远成[1] 蔡雨函[1] 朱玲[1,2] 徐志文[1,2] 郭万柱[1,2] 

机构地区：[1]四川农业大学动物生物技术中心,四川雅安620514 [2]四川农业大学动物疫病与人类健康四川省重点实验室,四川雅安620514

出　　处：《中国兽医学报》2014年第7期1039-1042,共4页Chinese Journal of Veterinary Science

基　　金：四川省科技支撑计划资助项目(2012NZ0001)

摘　　要：根据GenBank中猪环曲病毒N基因设计合成2对特异性扩增引物,成功建立了猪环曲病毒的套式RT-PCR快速检测方法。为验证本方法的特异性,以猪伪狂犬病病毒、猪传染性胃肠炎病毒、猪流行性腹泻病毒、A群猪轮状病毒、猪瘟病毒、猪嵴病毒、沙门菌及大肠杆菌作为阴性对照,结果显示只有在猪环曲病毒的核酸作为模板时才能扩增出预期的258bp和198bp 2条片段。敏感性试验显示,该方法能检测到的最低核酸含量为1pg。结果表明,建立的RT-PCR方法特异性强、灵敏度高,可用于猪环曲病毒的临床检测和流行病学调查。利用本方法对采集自四川省部分地区的50份临床样品进行检测,猪环曲病毒阳性率为30%。According to the sequence published by GenBank,two pairs of special primers were designed to amplify the N gene of porcine torovirus and a rapid nested RT-PCR mehtod for the detection of porcine torovirus has been developed.Two pairs of primers amplified 258bp and 198bp fragment respectively as expected.This method has good reproducibility,specificity and sensitivity,may detect low content of PToV accurately and rapidly.In a total of 50clinical samples collected from Sichuan province by using the method the results,pig torovirus positive rate was 30%in clinical samples.Ther result showed the nest RT-PCR mehtods would be used to detect porcine torovirus,which can support the clinic detection and molecular epidemiological survey.

关 键 词：猪环曲病毒 套式RT-PCR 检测 

分 类 号：S852.65[农业科学—基础兽医学]