黑龙江部分地区野鸟源新城疫病毒HN、P基因的克隆和序列分析

Cloning and sequence analysis of HN and P gene of Newcastle disease virus isolated from wild birds in some areas of Heilongjiang China

为了解野鸟源新城疫病毒(NDV)基因变异情况,我们对14株2005年～2006年分离自黑龙江三江自然保护区和哈尔滨周边地区健康野鸟体内的NDV的HN和P基因分别进行RT-PCR扩增,并将其克隆至pMD18-T载体进行核苷酸序列测定。结果显示:14株野鸟源NDVHN基因开放性阅读框架(ORF)均为1716bp,编码571个氨基酸;P基因开放阅读框均为1188bp,编码395个氨基酸。13株野鸟源NDVHN和P基因核苷酸同源性分别为96.4%～99.8%和95.8%～100%,分别与参考毒株HN和P基因相比,同源性为82.3%～98.7%和78.5%～99.4%。1株野鸟源NDV与参考毒株Mutkeswar的HN和P基因高度同源。分析表明:这14株NDV病毒与我国普遍流行的基因Ⅶ型和基因Ⅲ型NDV具有很高的同源性,提示野鸟与家禽新城疫的发生在流行病学和生态学上有着非常密切的关系。

The full-length HN and P genes of 14 Newcastle disease virus （NDV） isolates isolated from healthy wild birds in Heilongjiang province were amplified by RT-PCR,cloned and sequenced. The HN gene was 1 716 bp in length and contained a single opening reading frame （ORF） coding 571 amino acids,and P gene contained 1 188 bp coding 395 amino acids. The nucleotide homology of HN and P gene were 96.4 % to 99.8 % and 95.8 % to 100 % respectively among the 13 strains,and 82.3 % to 98.7 % and 78.5 % to 99.4 % respectively compared with these of the reference strain. One wild bird NDV strain is highly homologous with vaccine strain Mutkesear. These results indicated that the wild bird NDV strains prevalent in Heilongjiang are highly homologous with VIId subgenotype and Ⅲ subgenotype. These findings imply that the wild birds may be closely related to the Newcastle disease outbreaks in poultry.