贵州省地方品种鸡禽白血病病毒流行株的亚型鉴定及其分子特征分析

发布时间：2018-04-06 04:31

本文选题：禽白血病病毒　切入点：贵州　出处：《中国预防兽医学报》2017年03期

【摘要】：为了解贵州省9个地方品种鸡群中禽白血病病毒(ALV)流行株亚型与分子特征,本研究2016年从该省8个地区9个地方品种鸡群共采集380份鸡蛋样品。采集卵白用ELISA试剂盒检测ALV-P27抗原,阳性卵白接种CEF和DF1细胞分离病毒,培养7 d后检测细胞培养液中ALV-P27抗原,对感染细胞cDNA以4对ALV特异性引物进行PCR鉴定与分型。并选取9个鸡群的部分ALV分离株进行gp85基因的克隆测序与序列分析。结果表明,9个鸡群的卵白样品ALV-P27抗原阳性率为0~36%;阳性卵白的CEF和DF1细胞培养上清P27抗原检测结果相同,阳性率为26.8%(15/56),并鉴定出其亚型全部为J亚型(ALV-J)。已测序的21株ALV-J gp85基因序列与国内外ALV-J参考株同源性为86.6%~100%;进化树分析显示,21株ALV-J分离株在不同的分支,其进化关系和来源之间未发现明显规律。本研究表明,贵州省9个地方品种鸡群有6个存在ALV-J感染。其中,3个当地培育的品种鸡ALV感染率为0;而来自3个地区的绿壳蛋鸡ALV-P27抗原阳性率普遍较高(26.7%~36.0%)。本研究补充了我国禽白血病流行病学的数据并为该病的防制提供了调查依据。
[Abstract]:In order to understand the subtypes and molecular characteristics of avian leukemia virus (ALV) epidemic strains in 9 local breeds of chickens in Guizhou Province, 380 egg samples were collected from 9 local breeds of chickens in 8 regions of Guizhou province in 2016.Egg white was collected and tested for ALV-P27 antigen with ELISA kit. The positive egg white was inoculated with CEF and DF1 cells to isolate the virus. After 7 days of culture, the ALV-P27 antigen in the culture medium was detected. The cDNA of infected cells was identified and typed by PCR with 4 pairs of ALV specific primers.Partial ALV isolates from 9 chicken populations were selected for cloning, sequencing and sequencing of the gp85 gene.The results showed that the positive rate of ALV-P27 antigen in egg white samples of 9 flocks was 0 ~ 366.The positive rate of P27 antigen in the supernatant of CEF and DF1 cells cultured with positive egg white was the same, and the positive rate was 26.8%, and all the subtypes were identified as subtype J ALV-JN.The sequence of 21 ALV-J gp85 genes sequenced was 86.6% homology with ALV-J reference strains at home and abroad, and the phylogenetic tree analysis showed that 21 ALV-J isolates were in different branches, and there was no obvious regularity between their evolutionary relationships and sources.The results showed that ALV-J infection was found in 6 chickens of 9 local breeds in Guizhou province.Among them, the ALV infection rate of three local breeds was 0, while the positive rate of ALV-P27 antigen of green shell laying hens from three regions was generally higher than 26.7g / 36.0.This study supplements the epidemiological data of avian leukemia in China and provides an investigation basis for the prevention and control of the disease.
【作者单位】：山东农业大学动物科技学院;
【基金】：国家公益性行业(农业)科技专项(201203055) 江苏省动物预防医学重点实验室开放课题
【分类号】：S852.65

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