猪源副粘病毒的分子鉴定

Molecular identification of porcine paramyxovirus isolate

参照GenBank发表的相关猪源副粘病毒和禽源副粘病毒融合蛋白F基因核苷酸序列,设计并合成了4对引物,采用RT-PCR技术对本研究室分离的猪源副粘病毒JL-1株F基因进行探索性扩增、序列测定,在此基础上设计引物扩增HN基因、进行序列测定,分析F、HN基因序列,对该病毒进行分子鉴定。结果显示,扩增出了JL-1株F、HN基因目的条带,序列分析表明其与猪源副粘病毒病家族其他成员即蓝眼病病毒、尼帕病毒、梅那哥病毒同源性很低,与禽副粘病毒1型(APMV-1)具有高度同源性。初步确定分离的猪源副粘病毒为禽副粘病毒1型。

According to nucleotide sequence of porcine paramyxovims（PPMV ）and avian paramyxovims serotype 1 （APMV-1 ） in GenBank, four pairs of primers were designed and synthesized.The fusion protein F gene of a domestic isolate strain（PPMV JL- 1 strain）was amplified by RT-PCR,cloned into pMD18-T vector,sequenced and analyzed.Then the FIN was amplified by RT-PCR, cloned into pMD18-T vector, sequenced and analyzed. The results revealed that the F gene and HN gene of PPMV JL- 1 strain could be amplified by RT-PCR. Using the primers which designed depending on the nucleotide sequence of APMV-1. Compared with other members of PPMV（Nipah virus,Menangle virus,La Picdad Michoacan virus） ,the F gene and HN gene of PPMV JL-1 strain has a low homology. But has a high homology with APMV-1 .With the results we make sure that PPMV JL-1 strain belongs APMV-1.