建立用于检测马流感病毒核蛋白的抗原捕获 ELISA 方法
Development of an antigen-capture ELISA for the detection of equine influenza virus nucleoprotein.
机构信息
Division of Livestock Infectious Diseases, State Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences, 427 Ma Duan Street, Harbin 150001, China.
出版信息
J Virol Methods. 2011 Jul;175(1):120-4. doi: 10.1016/j.jviromet.2011.04.016. Epub 2011 Apr 28.
An antigen-capture enzyme-linked immunosorbent assay (AC-ELISA) was developed for the detection of the equine influenza virus (EIV), employing monoclonal and polyclonal antibodies against the A/equine/Xingjiang/2007 (H3N8) nucleoprotein (NP). Immunoglobulin G antibodies were purified and used as capture or detector antibodies. The specificity of the optimized AC-ELISA was evaluated using EIV, equine herpesvirus 1 (EHV-1), equine herpesvirus 4 (EHV-4), equine arteritis virus (EAV) and Japanese encephalitis virus (JEV), resulting in only EIV specimens yielding a strong signal. A minimal concentration of 50 ng/ml EIV protein was detected in Nonidet P40-treated virus preparations. Virus from the nasal swabs of equines infected experimentally were detected from days 3 to 7 post-infection using this AC-ELISA, with results confirmed by virus isolation and multi reverse transcription polymerase chain reaction. Both H3N8 and H7N7 EIV subtypes were AC-ELISA positive, indicating that this assay is suitable for the detection of all EIV subtypes.
建立了一种用于检测马流感病毒（EIV）的抗原捕获酶联免疫吸附试验（AC-ELISA），该试验使用针对 A/equine/Xingjiang/2007（H3N8）核蛋白（NP）的单克隆和多克隆抗体。免疫球蛋白 G 抗体被纯化并用作捕获或检测抗体。使用 EIV、马疱疹病毒 1（EHV-1）、马疱疹病毒 4（EHV-4）、马动脉炎病毒（EAV）和日本脑炎病毒（JEV）评估优化后的 AC-ELISA 的特异性，结果仅 EIV 标本产生强信号。在非离子型去污剂处理的病毒制剂中检测到 50ng/ml 的最小 EIV 蛋白浓度。使用该 AC-ELISA 从感染实验的马的鼻拭子中检测到感染后第 3 至 7 天的病毒，结果通过病毒分离和多重逆转录聚合酶链反应得到证实。H3N8 和 H7N7 EIV 亚型均为 AC-ELISA 阳性，表明该检测方法适用于所有 EIV 亚型的检测。
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