5株禽新城疫病毒新疆分离株F基因序列测定与遗传进化分析

The Genetic Sequence Detection and Analysis of F gene of Five Birds NDV Strains Isolated from Xinjiang

从新疆乌鲁木齐市郊区养禽场发病禽群中分离到4株鸡源新城疫病毒(Newcastle disease virus,NDV)和1株鸽源NDV。经血凝HA试验鉴定,NDV分离株均具有血凝性,5个毒株的血凝特性均可被NDV阳性血清所抑制,而不能被禽流感病毒(H5亚型与H9亚型)阳性血清抑制。本试验通过RT-PCR扩增了5株NDV全长F基因并进行了序列测定。序列分析表明,5株NDV的核苷酸序列分别为1662、1589、1676、1589和1662bp,均含有1个开放阅读框,分別编码553、528、553、520和553个氨基酸;根据基因裂解位点的氨基酸序列分析表明,鸡源XJ/10/08株属于强毒株,XJ/3/07株、XJ/7/07株、XJ/9/08株和XJ/11/08株属于弱毒株;同源性分析表明,5个NDV新疆分离株与LaSota疫苗株的核苷酸同源性在83.7%～99.8%之间,与TaiWan95株核苷酸同源性在84.7%～93.3%之间;遗传发育进化树分析表明,分离到的4个弱毒株与La-Sota、Clone30、B1、BEA-45在同一亚群,属基因Ⅱ型,强毒分离株与TaiWan95、广东株(GD-1-98)、江苏株(JS-5-1)在同一进化分支内,属基因Ⅶ型。

Five Newcastle disease virus strains were isolated from chickens and pigeon of poultry farm in Xinjiang. The resuits of the hemagglutination test （HA） and hemagglutination inhabition test （HI） indicated that the 5 NDV strains had HA. The HA of strains all can be inhibited by NDV positive serum and can＇t be inhibited by AIV （H5 subtype and H9 subtypes） positive serum. In this study, the F gene of strain XJ/3/07, strain XJ/7/07, strain XJ/9/08, strain XJ/10/08 and strain XJ/ 11/08 were amplified by RT-PCR and cloned. The sequence analysis showed that the full-length gene of F were 1662 bp, 1589 bp, 1676 bp, 1589 bp and.1662 bp and encoding 553,528,553,520 and 553 amino acids respectively. According to the analysis of amino acid sequence of the cleavage site of F protein, the NDV strain XJ/10/08 was virulent and the others NDV strains were avirulent. Comparison the five NDV strains with the vaccine strain La Sota and strain TaiWan95, the nucleotide ho- mology were 83.7% to 99. 8% and 84. 7% to 93. 3%. The phylogenetic analysis suggested that the strain XJ/3/07, strain XJ/7/07, strain XJ/9/08 and strain XJ/11/08 were gene type Ⅱ , and the strain XJ/10/08 strain was gene type Ⅶ.