中国畜牧兽医2012，Vol.39Issue(9)：38-42,5.
猪传染性胃肠炎与流行性腹泻病毒二重RT-PCR检测方法的建立及应用
Establishment and Application of Duplex RT-PCR Assay for Detection of Transmissible Gastroenteritis Virus and Porcine Epidemic Diarrhea Virus
摘要
Abstract
A duplex RT-PCR for detection porcine transmissible gastroenteritis virus (TGEV) and porcine epidemic diarrhea virus (PEDV) was developed using the primers designed basing on the N and M gene coding for the nucleoprotein of TGEV and membrane glycoprotein of PEDV respectively. The total RNA of standard TGEV and PEDV strains were used as the positive control to establish the duplex RT-PCR assay. The sensitivity, specificity and repetition assay of duplex RT-PCR were tested, and samples taken from clinic suspicious TGEV or PEDV infected pigs and having been testified by routine cell culture and identification were detected by the established duplex RT-PCR assay. The results indicated that the duplex RT-PCR assay was successfully established. The specificity and sensitivity of the duplex RT-PCR assay revealed that the threshold of duplex RT-PCR was 10 TCID50/mL of TGEV or PEDV, and no products were amplified from the nucleic acid of ST cell or the other 7 kinds of pathogenic viral or bacterial microorganism acting as the negative control. The repetition test indicated that the duplex PCR was repeatable. The detection results for 22 clinic suspicious TGEV or PEDV infected pigs were consistent with the results tested by sequencing. The study suggested that the established duplex RT-PCR method was highly specific and sensitive, and was suitable to clinic rapid identified diagnosing of TGEV and PEDV.关键词
猪传染性胃肠炎病毒/猪流行性腹泻病毒/二重RT-PCR/检测/建立/应用Key words
porcine transmissible gastroenteritis virus/ porcine epidemic diarrhoea virus/ duplex RT-PCR/ detection establishment / application分类
农业科技引用本文复制引用
闫若潜,安春霞,刘淑敏,赵雪丽,郭小玲,赵明军,吴志明..猪传染性胃肠炎与流行性腹泻病毒二重RT-PCR检测方法的建立及应用[J].中国畜牧兽医,2012,39(9):38-42,5.基金项目
河南省科技攻关资助项目(0524030003). （0524030003）
