种禽场A亚群禽白血病病原学调查及分离株遗传进化分析  

Etiology surveys and phylogenetic analyses of avian leukosis virus subgroup A isolated from breeder chickens farms

作　　者：冯敏[1] 谭利强 代曼曼[1] 郝建勇[1] 秦建如 黄小容[1] 廖明[1] 曹伟胜[1] 

机构地区：[1]华南农业大学兽医学院/农业部兽用疫苗创制重点实验室,广东广州510642

出　　处：《华南农业大学学报》2014年第4期11-15,共5页Journal of South China Agricultural University

基　　金：农业部公益性行业科研专项(201203055);广东省自然科学基金(S20110100001946);国家肉鸡产业技术体系(nycytx-42-G3-03);广东省科技计划项目(2012A020100001)

摘　　要：【目的】了解广东地区种禽场A亚群禽白血病病毒(ALV-A)流行情况.【方法】从A、B、C、D 4个种禽场采集1 561份血浆样品,接种DF-1细胞,培养7 d后通过ELISA方法对细胞上清液进行p27抗原检测,通过PCR、间接免疫荧光试验2种方法对p27抗原阳性样品进行鉴定.【结果和结论】从4个种禽场共检出71份阳性样品,外源性禽白血病病毒分离阳性率为4.6%.其中,仅从C种禽场分离到2株ALV-A,命名为GD13-1和GD13-2,前病毒全基因序列分别为7 721和7 715 bp,且GD13-2与J亚群禽白血病病毒混合感染.与国内外其他ALV-A的LTR、gp85核苷酸序列进行分析比对,发现该研究分离的2株ALV-A与国内A亚群分离株SDAU09E2相似度最高,其中与LTR相似性分别为96.9%、97.2%,与gp85相似性分别为98.4%、98.7%;与广东地区5年前ALV-A分离株GD08的LTR核苷酸序列相似性分别为88.9%、89.5%,与gp85相似性分别为98.5%、98.8%.调查结果表明,广东地区部分种禽场内仍然存在ALV-A感染,但ALV-A已经不是流行毒株,且变异程度不大.Objective]To investigate the prevalence of breeders infected by avian leukosis virus subgroup A （ ALV-A） in Guangdong Province of China .[Method] A total of 1 561 plasma samples of breeders collected from four different farms （ A,B,C,D） were inoculated into DF-1 cell cultures for 7 days; the positive samples were detected by antigen p 27 ELISA test and identified via PCR and IFA .[Result and conclusion]Results showed that 71 positive samples were detected from four farms and the positive rate of exogenous ALV isolation was 4 .6%.Two ALV-A viruses were isolated from antigen p 27 ELISA positive samples of the breeder chickens farm C , which were designed as GD13-1 and GD13-2, and the GD13-2 was determined for mixed infection with ALV-J.The full length proviral genomes of two isolates were 7 721 , 7 715 bp after PCR amplification and sequencing .In comparison to the LTR and gp85 sequence of the other ALV-A strains at home and abroad , the isolates had the highest similarity with that of sub-group A strain SDAU09E2.The nucleotide similarity of LTR and gp85 were 96.9%, 97.2% and 98.4%, 98.7%respectively .In comparison to the LTR and gp85 sequence of ALV-A GD08 isolated from Guangdong five years ago , the similarity of LTR and gp85 were 88.9%, 89.5% and 98.5%, 98.8%respectively .The survey results show that ALV-A still exist in some breeder farms , but it is not prevalent any more .

关 键 词：种禽场 A亚群禽白血病病毒 前病毒全基因组序列 

分 类 号：S855.3[农业科学—临床兽医学]