【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 p27 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 p27 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 subgroup 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.
冯敏,谭利强,代曼曼,郝建勇,秦建如,黄小容,廖明,曹伟胜.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[J].华南农业大学学报,2014,35(4):11-15复制