猪细环病毒两种基因型双重PCR检测方法的建立

Development of duplex PCR for detection of both porcine Torque Teno virus genogroups

为建立检测猪细环病毒两种基因型(PTTV1和PTTV2)的双重PCR方法,本研究根据GenBank中PTTV1、PTTV2的UTR基因序列,设计合成了2对特异引物,并通过对扩增条件的筛选,建立了PTTV1和PTTV2的双重PCR检测方法。该方法可以同时扩增PTTV1的324 bp和PTTV2的522 bp特异性片段,而扩增猪圆环病毒2型和猪细小病毒DNA结果均为阴性,对PTTV1和PTTV2的最低检出量分别为100 copies和10 copies。该方法适合对PTTV1和PTTV2的联合检测。

To develop a rapid differential detection of both porcine Torque Teno virus genogroups （PTTV1 and PTTV2）, a duplex PCR was successfully developed with two pairs of special primers designed based on the untranslated region sequences of PTTV1 and PTTV2, respectively, under the optimized amplification conditions, the specific PCR products of 324 bp for PTTV1 and 522 bp for PTTV2 were simultaneously amplified in the duplex PCR, whereas no PCR products were amplified from porcine circovirus type 2 and porcine parvovirus. The duplex PCR method was capable of detecting the template DNA of 100 copies for PTTV1 and 10 copies for PTTV2, and was able to simultaneously detect for PTTV1 and PTTV2 in a mixed infections.