RT-PCR技术检测猪流感病毒

根据猪流感病毒（SIV）的M基因的序列，设计合成了1对引物，建立了检测猪流感的RT-PCR方法。应用该方法对H1、H3、H9亚型的猪流感病毒进行基因扩增，均获得了分子量为460bp的特异性目的片段，而对PRRSV、 PCV2、PRV、CSFV进行同条件检测，结果均为阴性；SIV扩增产物测序结果与SIV其他毒株序列同源性达99%～100% 。敏感性试验表明，该方法可检测到103 EID50的SIV；可直接从猪流感病毒感染小鼠的组织样品中检测到病毒。

RT-PCR detection of Swine influenza virus

Based on the Mtrix（M）gene sequence of the Swine influenza virus(SIV),a pair of primers were designed and synthesized. By means of it, the RT-PCR technique for detecting SIV was established. The 460 bp specific fragments were obtained subsequently from the Mtrix（M）gene of H1,H3 and H9 subtype Swine influenza viruses. Negative test results were found on the Porcine reproductive and respiratory syndrome virus，Porcine circovirus type 2, Swine pseudorabies virus and Classical swine fever virus. The amplified sequence of PCR product was 99%-100% in agreement with those published data on SIV strains. The sensitivity of RT-PCR reached 103 EID50. The Swine influenza virus could be detected directly from the samples of infected mice ’s lungs.