应用荧光定量PCR技术快速定量检测猪瘟病毒

Rapid Quantitative Detection of Classical Swine Fever Virus by Real-time TaqMan Assay

采用荧光定量PCR技术对猪瘟病毒进行快速定量检测.实验证明,该方法的检测灵敏度可达10拷贝/μL,对于不同毒株以及各种组织样品检测结果均为阳性.通过该方法与RT PCR、nPCR的比较,发现该方法检测灵敏度比RT PCR高100倍,与nPCR具有相同的灵敏度,并且该方法避免了常规PCR电泳检测所带来的高污染率.因此,该方法以其快速、灵敏、低污染率的优点,会在猪瘟的早期检测及预防、控制上起到重要作用.

A real-time TaqMan method was developed for detection of classical swine fever virus(CSFV).According to our results, CSFV was detected specifically from all kinds of tissues and blood specimens from pigs experimentally infected with high virulent CSFV,and moderate、low virulent strains of CSFV respectively. It has a sensitivity of 10 copies/μL.By serial dilution of CSFV samples, the TaqMan method was found to 100-fold times more sensitive than RT\|PCR, and it can avoid contamination with non-specific amplicons by post-PCR analysis. Therefore, it was sensitive、specific and rapid for early diagnose of CSFV-related infection.