Production of monoclonal antibody against ORF72 of koi herpesvirus isolated in Taiwan
Production of monoclonal antibody against ORF72 of koi herpesvirus isolated in Taiwan
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DOI：
10.1007/s12223-013-0261-7
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发表时间：
2013-09
影响因子：
2.6
通讯作者：
C. Tu;Yi-Ping Lu;C. Hsieh;Su-Ming Huang;Shao-Kuang Chang;Meei-Mei Chen
中科院分区：
文献类型：
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作者：
C. Tu;Yi-Ping Lu;C. Hsieh;Su-Ming Huang;Shao-Kuang Chang;Meei-Mei Chen
从锦鲤（koi Cyprinus carpioin Taiwan）分离到锦鲤疱疹病毒（koi herpesvirus，KHV），并制备了KHV衣壳蛋白（ORF 72）的单克隆抗体。用全病毒颗粒免疫小鼠，获得MAb-B2克隆，用间接酶联免疫吸附试验（ELISA）和Western blot鉴定。此外，它检测到KHV感染的细胞，但不是在那些模拟感染的细胞中的KHV的间接免疫荧光试验证明。中和试验表明MAb-B2对KHV具有中和作用。此外，我们发现，MAb-B2识别的ORF 72的KHV所揭示的液相色谱-串联质谱和蛋白质印迹分析。此外，MAb-B2已被用作通过免疫组织化学检测临床样品中KHV的诊断工具。本研究结果表明，单克隆抗体B2可用于KHV感染诊断试剂盒的研制，KHV ORF 72蛋白可作为KHV疫苗的候选抗原。
A monoclonal antibody (MAb) was generated against the capsid protein (ORF 72) of koi herpesvirus (KHV) isolated from diseased koiCyprinus carpioin Taiwan. The clone of MAb-B2 was obtained by immunizing mice with whole virus particles and further identified using indirect enzyme-linked immunosorbent assay and Western blot assay. In addition, it detected KHV in KHV-infected cells but not in those of mock-infected cells as demonstrated by indirect immunofluorescence assay. The neutralization test showed that MAb-B2 neutralized KHV. Furthermore, we uncovered that MAb-B2 recognizes the ORF72 of KHV as revealed by liquid chromatography–tandem mass spectrometry and Western blot assays. Additionally, MAb-B2 has been used as a diagnostic tool for detection of KHV in clinical samples by immunohistochemistry. Collectively, our results indicated that MAb-B2 could be used in the development of a diagnostic kit for diagnosis of KHV infections and ORF72 protein of KHV might be a candidate for future vaccine development.