半番鸭胚中鹅细小病毒的分离鉴定及其基因组分子特征分析

摘要

A goose parvovirus (GPV) strain (named MDE) was isolated from mule duck embryos and mule ducks hatched and identified by colloidal gold labeled technique,electron microscope and PCR.The results shows that the virions were spherical with a diameter of approximately 20-24 nm.The colloidal gold test results were that GPV is positive.The results of complete genomic sequences analysis demonstrated that the full-length genome of MDE strain is 5106 bp in length.The nucleotide homology of genomic sequences between MDE strain and other 15 GPV isolates ranged from 93.2 ％ to 98.1 ％,sharing the greater similarity to strain SH and the lower similarity to strain M15 isolated from mule ducks.Also,the VP1 gene shared higher similarity to the vaccine SYG61v strain.Phylogenetic analysis of VP1 and NS1 gene showed that MDE was a distinct lineage of goose parvovirus but separate from Muscovy duck parvovirus (MDPV).Compared with the vaccine SYG61v strain,MDE add the deduced 703NRTS glycosylation in VP1 protein.In conclusion,The isolated GPV strains in this study can be transmitted vertically by mule duck embryos and enriched the ecology of GPV.%通过胶体金试验、电镜观察及PCR扩增等方法,自半番鸭胚及孵化的雏半番鸭中分离获得了鹅细小病毒(GPV)毒株(命名为MDE株),并进行了病毒全基因组分子特征分析.结果表明,该病毒粒子为直径20～24 nm的球形、无囊膜粒子.胶体金试验结果呈GPV阳性.该毒株基因组全长为5 106 bp,与GenBank登录的15株GPV基因组序列相似性为93.2％～98.1％,其中与SH株及疫苗株SYG61v株等的相似性较高,而与半番鸭源的GPV重组病毒M15株相似性最低.基于病毒VP1和NS1基因的遗传进化分析表明,分离毒株MDE与GPV疫苗毒株的亲缘关系最近,处于同一个独立的分支上,与番鸭细小病毒的亲缘关系较远.与疫苗毒株SYG61v株比较,MDE株VP1区的糖基化位点增加了703NRTS糖基化位点.以上结果表明,本研究揭示了GPV可通过半番鸭胚垂直传播给雏鸭,丰富了GPV的生态学内容.