摘要
本研究根据猪传染性胃肠炎病毒(TGEV)N基因和猪肌动蛋白的基因序列设计合成了引物和探针,RT-PCR扩增得到目的基因,并克隆到pMD18-T载体得到阳性质粒,即为质粒标准品。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。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。
In the study, the primers and probes were designed and synthesized according to the sequences of porcine transmissible gastroenteritis virus and 13- actin. The genes were amplified by reverse transcription polymerase chain reaction （ RT - PCR） and cloned to the pMD18 - T vector. Then we got the standards. The reaction parameters were optimized to develop TaqMan fluorescence quantitative RT- PCR assay. It was showed that the fluores, cence quantitative RT - PCR assay could detect 15.3copies/μL of plasmid DNA and its Specificity and reproducibility were very good.
出处
《检验检疫科学》
2009年第5期17-20,共4页
Inspection and Quarantine Science
