猪传染性胃肠炎病毒Taq Man荧光定量RT-PCR检测方法的建立  

Development of TaqMan Fluorescence Quantitative RT-PCR Assay for Detection of Transmissible Gastroenteritis Virus of Swine

作　　者：白兴华[1] 冯力[1] 陈建飞[1] 时洪艳[1] 孙东波[1] 吴波平[1] 高秀春[1] 

机构地区：[1]中国农业科学院哈尔滨兽医研究所兽医生物技术国家重点实验室猪传染病研究室,哈尔滨150001

出　　处：《畜牧兽医学报》2007年第5期476-481,共6页ACTA VETERINARIA ET ZOOTECHNICA SINICA

摘　　要：根据猪传染性胃肠炎病毒和猪肌动蛋白的基因序列设计合成了引物和探针,通过对荧光定量RT-PCR反应条件的优化,建立了TaqMan荧光定量RT-PCR检测TGEV的方法。同时对37份现地病料进行检测并与常规RT-PCR方法、TGEV抗原快速检测试剂盒比较。结果,该方法的检测敏感性达到15.3拷贝/μL,且具有很好的特异性和重复性,而常规RT-PCR方法只能检测到1.53×103拷贝/μL。对37份现地病料的检测结果也表明该法(检出17份)比常规RT-PCR方法(检出12份)和TGEV抗原快速检测试剂盒(免疫层析法,检出10份)的敏感性高。The primers and probes were designed and synthesized according to the sequences of transmissible gastroenteritis virus and β-actin, and then reaction requirements were optimized to develop a TaqMan fluorescence quantitative RT-PCR assay. Meanwhile,37 field samples were detected and the results were compared with that of routine RT-PCR and antigen rapid test kit. It was showed that the fluorescence quantitative RT-PCR assay could detect 15.3 copies/μL of plasmid DNA and its specificity and reproducibility were very good, while the sensitivity of the routine RT-PCR was 1.53 × 10^3 copies/μL. The results of field test also showed that its sensitivity was higher than that of the routine RT-PCR and TGEV antigen rapid test kit （chromatographic immunoassay）.

关 键 词：猪传染性胃肠炎病毒 TAQMAN探针 荧光定量RT-PCR 

分 类 号：S852.65[农业科学—基础兽医学] S854.43[农业科学—兽医学]