猪圆环病毒2型TaqMan实时PCR检测方法的建立

Development of a Real-time PCR Method for Detection of Type 2 Porcine Circovirus

设计合成了一套引物和TaqMan探针,特异性扩增猪圆环病毒2型(PCV2)ORF2基因,在国内首次建立了快速定量检测PCV2的实时PCR方法,且该方法具有较好的特异性和重复性,对PCV2DNA检测下限为1copy/μL,敏感性比常规PCR高106倍;分别用该法和普通PCR方法对PMWS人工发病猪的10份组织及30份血清样品检测,结果表明该方法具有更快速、灵敏、准确、低污染等优点,并可以对PMWS的早期检测、预防起到指示作用。

One set of primers and a TaqMan probe were designed based on the nucleotide sequence within ORF2 of Porcinecircovirus type 2 （PCV2）. A quantitative TaqMan real-time PCR was developed for detecting PCV2 with highly specificity, reproducibility and more sensitivity than conventional PCR. Standard dilutions allowed absolute quantification of the amount of viral DNA to Icopy/μL, which is 10^6 times more sensitive than conventional PCR. The result of detecting PCV2 from tissues and sera of pigs with PMWS also confirms the high sensitivity and specificity of this method. This method is probably a powerful enough tool for early diagnosis of PMWS.