Tadese Theodros, Reed Willie M
Department of Pathobiology and Diagnostic Investigation, Diagnostic Center for Population and Animal Health, Michigan State University, A510 East Fee Hall, East Lansing, MI 48824, USA.
J Virol Methods. 2003 Jun 9;110(1):99-104. doi: 10.1016/s0166-0934(03)00106-x.
The detection is described of reticuloendotheliosis virus (REV) protein in tissue culture of chicken embryonated cells (CEFs) infected with field isolates of fowl poxvirus (FPV). By the polymerase chain reaction (PCR), five out of the six field isolates, but two out of the seven vaccine strains of FPV, were found to have had a 291 bp repeat sequence of REV-LTR integrated in their genomic DNA. An immunofluorescence (IF) method was employed using a monoclonal antibody (MAb) known to specify strain common envelope proteins for REV and allowed to detect the presence of a specific REV protein. The IF results indicate the localization of REV proteins in boundaries defined precisely within cells infected with these field strains of FPV carrying REV (FPV-REV). Furthermore, by immunoblotting (IB) using a chemiluminescent detection kit, the REV protein reacted specifically with the MAb and had a relative molecular mass (RMM) of 62 kDa. The data have the potential to advance substantially the current understanding of the integrated REV in FPV strains; and the identification of a unique protein associated with variant forms of FPV will also offer great potential for identification of novel vaccine candidates for use in poultry against variant forms of FPV.
描述了在感染禽痘病毒(FPV)野外分离株的鸡胚成纤维细胞(CEF)组织培养物中检测网状内皮组织增生症病毒(REV)蛋白的情况。通过聚合酶链反应(PCR)发现,6株野外分离株中有5株,但7株FPV疫苗株中有2株,其基因组DNA中整合有REV-LTR的291 bp重复序列。采用免疫荧光(IF)方法,使用已知能特异性识别REV株共同包膜蛋白的单克隆抗体(MAb),检测特定REV蛋白的存在。IF结果表明,REV蛋白定位于感染携带REV的这些FPV野外株(FPV-REV)的细胞内精确定义的边界处。此外,通过使用化学发光检测试剂盒的免疫印迹(IB),REV蛋白与MAb发生特异性反应,相对分子质量(RMM)为62 kDa。这些数据有可能极大地推进目前对FPV株中整合的REV的理解;并且鉴定与FPV变异形式相关的独特蛋白也将为鉴定用于家禽对抗FPV变异形式的新型候选疫苗提供巨大潜力。