摘要
据猪传染性胃肠炎病毒(transmissible gastroenteritis virus of swine,TGEV)S基因序列设计1对特异性引物,通过对实时荧光定量RT-PCR反应条件的优化,建立了SYBR GreenⅠ实时荧光定量RT-PCR检测TGEV的方法,同时对12份病料进行检测,并与常规RT-PCR进行比较。结果显示,该方法的敏感性达到43.07拷贝/μL,具有良好的特异性和重复性,而常规RT-PCR最低只检测到4.307×103拷贝/μL,敏感性较低。本试验建立的检测TGEVS基因的SYBR GreenⅠ实时荧光定量RT-PCR方法为传染性胃肠炎的鉴别诊断及TGEV的分离鉴定奠定了技术基础。
The primers were designed and synthesized according to the sequences of S gene of transmissible gastroenteritis virus （TGEV）, and then reaction requirements were optimized to develop a SYBR Green Ⅰ fluorescence quantitative RT-PCR assay. Meanwhile, 12 field samples were detected and the results were compared with that of routine RT-PCR. It was showed that the fluorescence quantitative RT-PCR assay could detect 43.07 copies/μL of plasmid DNA and its specificity and reproducibility were very good, while the sensitivity of the routine RT-PCR was 4.307×103 copies/μL.The results of field test also showed that its sensitivity was higher than that of routine RT-PCR. The SYBR Green Ⅰ fluorescent quantitative RT-PCR assay for detecting S gene of TGEV was developed for the basis of early， rapid detection and quantitative analysis of the infection intensity of TGEV in this experiment.
出处
《中国畜牧兽医》
CAS
北大核心
2014年第1期66-71,共6页
China Animal Husbandry & Veterinary Medicine
基金
山东省自主创新成果转化重大专项(2010ZHZX1A0417)
