麻黄肉种鸡蛋清中内源性禽白血病病毒的分离与遗传鉴定

Isolation and Genetic Identification of Endogenous Avian Leukosis Virus Strain from Albumen of Partridge breeder Chicken

为了解麻黄肉种鸡蛋清中内源性禽白血病病毒(内源性ALV)的分布情况,从广东某麻黄肉种鸡场采集1 038份蛋清,经ALV p27抗原ELISA检测出30份阳性蛋清;将阳性蛋清同时接种DF1和CEF细胞,培养7 d后,p27抗原ELI-SA检测到13份样品为DF1-CEF+;以这13份CEF+细胞培养物的基因组DNA为模板,经PCR扩增克隆测序最终得到了1株内源性ALV前病毒序列。将经ELISA、PCR和IFA等方法鉴定的该内源性ALV命名为HN1301株。基于遗传分析表明,HN1301株的前病毒基因序列与已知的E亚群毒株ev1、SD0501的相似性均为99.4%。表明该麻黄肉种鸡品系中存在有复制能力的内源性ALV,因而易干扰基于p27抗原的ELISA检测;本研究对于麻黄肉种鸡禽白血病的防控与净化有参考价值。

In order to investigate the distribution of endogenous avian leukosis virus in albumen of partridge breeders, 1 038 albumen were collected from one partridge breeder farm in Guangdong province, 30 positive samples were detected by antigen p27 ELISA test ; these positive albumen were inoculated into DF1 and CEF cell respectively and then cultivated for 7 days. By antigen p27 ELISA of celt cultures, 13 samples were DFl-and CEF＋. One whole genome of ALV strain was amplified, cloned and sequenced from the genomic DNA of CEF＋.The endogenous ALV isolate, determined as HN1301, was identified by methods of ELI- SA, PCR and IFA. Genetic analysis showed that HN1301 has the highest similarities with those of representative endogenous ALV strains evl and SD0501 based on provirus sequence. This study suggested there were endogenous ALV replications in partridge broiler chicken, which could interfere with ELISA if only based on p27 antigen detection .The study provides important information for the control and eradication of exogenous ALV from partridge broiler breeders.