多重PCR方法快速鉴别猪伪狂犬疫苗毒和野毒

Differentiation of Vaccine Strain and Field Isolate of Porcine Pseudorabies Virus by Developed Multiplex PCR

根据Genbank中PRV的gB、gE、TK基因序列设计并合成引物,对样品中PRVDNA进行多重PCR扩增及反应条件优化,得到与设计相符合的3条特异性条带,分别为427 bp(gB)、298 bp(gE)和208 bp(TK)。用这3对引物对四种伪狂犬疫苗样品DNA进行多次多重PCR扩增,其中1种疫苗得到与设计相符的1条特异性条带,其余为2条。该结果表明,此检测方法敏感度较高,适用于科学研究、临床诊断以及流行病学调查,对根除伪狂犬病具有重要意义。

According to the sequence of PRV gB, gE, and TK gene published in GenBank, the primers were designed. Multiplex PCR （multi-PGR） was developed and optimized to detect porcine pseudorbies virus, and the result revealed three specific bands at the expected size, respectively at 427 bp （gB gene）, 298 bp （gE gene）, and 208 bp （ TK gene）. Then four kinds of vaccine against pseudombies virus were detected by the developed multi-PCR. One expected specific band appeared in one sample, while two bands in the others. The detection results showed that the muhi-PCR has high sensitivity and should be applied to research, clinical diagnosis, and epidemiologieal investigation.