U.S. Department of Agriculture, Agriculture Research Service, Avian Disease and Oncology Laboratory, East Lansing, MI, USA.
Avian Pathol. 2010 Oct;39(5):383-9. doi: 10.1080/03079457.2010.510828.
Nine reticuloendotheliosis virus (REV) isolates obtained from broiler breeders, turkeys, and prairie chickens located in three different geographical regions in the USA, and three isolates obtained from known contaminated live-virus vaccines were characterized using polymerase chain reaction (PCR) and indirect immunofluorescence (IFA) assays. All isolates were propagated in chicken embryo fibroblasts obtained from a specific pathogen free breeder flock. PCR analysis of all 12 isolates resulted in the amplification of the 291-bp REV long-terminal repeat region (LTR); none of the isolates exhibited a different pattern or shift from the expected PCR product of REV LTR. The subtype of the REV isolates was determined by IFA using REV-specific monoclonal antibodies, 11B118.22, 11C237.8, and 11D182. Results from sub-typing indicated that all nine isolates from broiler breeders, turkeys, and prairie chickens belonged to subtype 3, and are antigenically related to the chick syncytial virus (CSV) strain of REV, the prototype of subtype 3 REV. In contrast, the three isolates from contaminated vaccines were classified as subtype 2, and were antigenically related to spleen necrosis virus (SNV) strain of REV, the prototype of subtype 2 REV. Three isolates representing REV isolated from broiler breeders, turkeys, and prairie chickens were cloned and further evaluated by DNA sequence analysis of the envelope gene. Results from DNA sequence analysis confirmed those from sub-typing and indicated that the three REV isolates representing those from broiler breeders, turkeys, and prairie chickens are closely related to CSV of REV, with an amino acid homology of 98% or greater as compared with SNV with an amino acid homology of 95% or less. Data from this study clearly indicate that subtype 3 is the most common subtype of REV circulating in three different avian species, namely broiler breeders, turkeys and prairie chickens, located in three different geographical regions in the United States.
从美国三个不同地理位置的肉鸡种鸡、火鸡和草原榛鸡中分离到的 9 株网状内皮组织增生症病毒（REV）和从已知污染活病毒疫苗中分离到的 3 株病毒，采用聚合酶链反应（PCR）和间接免疫荧光（IFA）方法进行了鉴定。所有分离株均在来自无特定病原体种鸡群的鸡胚成纤维细胞中繁殖。对所有 12 株分离株的 PCR 分析导致扩增出 291bp 的 REV 长末端重复区（LTR）；没有一株分离株的 PCR 产物与 REV LTR 的预期产物出现不同的模式或偏移。使用 REV 特异性单克隆抗体 11B118.22、11C237.8 和 11D182 通过 IFA 对 REV 分离株的亚型进行了测定。亚型测定结果表明，来自肉鸡种鸡、火鸡和草原榛鸡的 9 株分离株均属于 3 型，与 REV 的鸡合胞体病毒（CSV）株具有抗原相关性，CSV 是 3 型 REV 的原型株。相比之下，来自污染疫苗的 3 株分离株被归类为 2 型，与 REV 的脾坏死病毒（SNV）株具有抗原相关性，SNV 是 2 型 REV 的原型株。从肉鸡种鸡、火鸡和草原榛鸡中分离到的 3 株 REV 分离株进行了克隆，并通过 envelope 基因的 DNA 序列分析进一步进行了评估。DNA 序列分析结果证实了亚型分析结果，并表明代表肉鸡种鸡、火鸡和草原榛鸡的 3 株 REV 分离株与 REV 的 CSV 密切相关，与 SNV 的氨基酸同源性为 95%或更低，而与 CSV 的氨基酸同源性为 98%或更高。本研究的数据清楚地表明，3 型是在美国三个不同地理位置的三种不同禽类（肉鸡种鸡、火鸡和草原榛鸡）中循环的最常见的 REV 亚型。
