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首页> 外文期刊>Aquaculture >Development of monoclonal antibodies against VP28 of WSSV and itsapplication to detect WSSV using immunocomb
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Development of monoclonal antibodies against VP28 of WSSV and itsapplication to detect WSSV using immunocomb

机译：WSSV VP28单克隆抗体的开发及其在免疫梳检测WSSV中的应用

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White spot disease (WSD) is an important viral disease of penaeid shrimp caused by white spot syndrome virus (WSSV). WSSV isolated from WSD outbreaks in commercial shrimp (Penaeus monodon) farms in India were propagated in the laboratory in healthy shrimp. The virus was purified from the infected tissues by sucrose gradient centrifugation. The VP28 was electroeluted from SDS-PAGE gels and was used to immunize Balb/c mice to produce hybridomas secreting monoclonal antibodies (MAb) against WSSV. A total of five hybridoma clones secreting MAbs to VP28 were produced. The MAbs were of the isotypes IgG1, IgG2b and IgM. The MAbs reacted with VP28 of WSSV and not with any other viral or shrimp protein in western blot. The MAbs were used to develop dot immunoblot assay using an immunocomb to detect WSSV from field samples. The test developed had an analytical sensitivity of 625 pg and a diagnostic sensitivity of 100% compared to single step polymerase chain reaction (PCR). The test can be used as an alternate for first step PCR to detect WSSV from field samples.
机译：白斑病（WSD）是由白斑综合症病毒（WSSV）引起的对虾重要的病毒性疾病。从印度的商业对虾（对虾）养殖场的水产养殖爆发中分离出的WSSV在实验室中以健康对虾繁殖。通过蔗糖梯度离心从感染的组织中纯化病毒。 VP28从SDS-PAGE凝胶上电洗脱，用于免疫Balb / c小鼠以产生分泌抗WSSV单克隆抗体（MAb）的杂交瘤。总共产生了分泌MAb至VP28的五个杂交瘤克隆。 MAb具有同种型IgG1，IgG2b和IgM。免疫印迹中，单克隆抗体与WSSV的VP28反应，而不与任何其他病毒或虾蛋白反应。 MAb用于开发点免疫印迹测定法，使用免疫梳检测田间样品中的WSSV。与单步聚合酶链反应（PCR）相比，开发的测试的分析灵敏度为625 pg，诊断灵敏度为100％。该测试可以用作第一步PCR的替代方法，以从现场样品中检测WSSV。

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