云南省新城疫病毒分离株F基因和HN基因克隆与序列分析

Cloning and Sequence Analysis of F Gene and HN Gene from Newcastle Disease Virus Isolates in Yunnan

为了确定云南NDV毒株F基因和HN基因的结构特征及其与已知毒株的遗传相关性,应用RT-PCR技术对云南省部分地州养殖场采集的鸡或鸭喉气管组织样品4 700份进行新城疫病毒(NDV)检测,共检出阳性样品25份。对其中8份阳性样品进行NDV F基因与HN基因扩增后,克隆至pMD18-T载体测序,并与国内外代表性毒株和当前流行疫苗毒株进行比对及系统发育分析。结果表明,云南省NDV毒株F基因与国内代表性毒株核苷酸序列同源性介于93.0%～99.2%之间,氨基酸同源性介于90.7%～99.1%之间;与疫苗毒株的核苷酸同源性在83.3%～86.9%之间,氨基酸同源性在86.0%～88.7%之间。HN基因与国内代表性毒株核苷酸同源性介于96.4%～99.1%之间,氨基酸同源性介于95.6%～99.3%之间;而与当前疫苗毒株相比,同源性较低,核苷酸同源性介于81.3%～82.0%之间,氨基酸同源性介于87.6%～88.8%之间。系统进化分析表明,云南NDV F基因有2株属于基因Ⅱ型,其余6株均属于基因Ⅶe型。云南NDV HN基因则均属于基因Ⅶ型。

To determine the structural and genetic characteristics of NDV F and HN genes of Yunnan, the infection status of Newcastle disease virus was detected by the reverse transcriptase-polymerase chain reaction （RT-PCR） from 4700 tissue samples collected from Yunnan province, the positive samples were 25. The F and HN genes were amplified from 8 samples by using specific primers, purified and cloned into pMD18-T vector for sequencing. The results of sequence alignment and phylogenetic analysis showed that the nucleotide homology of F gene is 93.0%-99.2%, and amino acid homology is 90.7%-99.1% compared with represented strains. Compared with the current vaccine strains, the nucleotide homology is 83.3%- 86.9%, the amino acid homology is 86.0%-88.7%. The HN gene nucleotide homology is 96.4%-99.1%, the amino acid homology is 95.6%-99.3% compared with represented strains. Compared with the current vaccine strains, the nucleotide homology is 81.3%-89.0%, the amino acid homology is 87.6%-88.8%. The phylogenetic analysis showed that two of eight strains of Yunnan NDV F gene belong to II type, the other six strains belong to Ⅶ e type, while the Yunnan NDV HN gene belongs to VII type.