在果蝇细胞中表达的牛病毒性腹泻病毒（BVDV）截短E2蛋白：一种用于BVDV酶联免疫吸附测定（ELISA）的候选抗原。
Truncated E2 of bovine viral diarrhea virus (BVDV) expressed in Drosophila melanogaster cells: a candidate antigen for a BVDV ELISA.
作者信息
Marzocca M P, Seki C, Giambiagi S M, Robiolo B, Schauer R, Dus Santos M J, Scodeller E A, La Torre J L, Wigdorovitz A, Grigera P R
机构信息
Fundación de estudios en Virología Animal (FEVAN), Guamini 1682, Ciudad de Buenos Aires (C1440ESD), Argentina.
出版信息
J Virol Methods. 2007 Sep;144(1-2):49-56. doi: 10.1016/j.jviromet.2007.03.023. Epub 2007 May 21.
A simple and reliable indirect enzyme-linked immunosorbent assay for detection of antibodies directed against a major bovine viral diarrhea virus (BVDV) immunogen, the E2 glycoprotein (tE2-ELISA), has been developed using the recombinant C-terminal truncated E2 glycoprotein (tE2) expressed in a Drosophila melanogaster system. This strategy demonstrated that tE2 is secreted efficiently in the supernatant, no purification steps are necessary, it is easy to produce and carries out the post translational modifications necessary to preserve its native conformation. Preliminary analysis of 183 cattle serum samples using tE2-ELISA showed a 98% specificity and a 100% sensitivity compared with the standard homologous BVDV virus neutralization test. The results also showed that the tE2 is immunoreactive because the conformation and antigenicity of the original E2 are maintained to a large extent. To our knowledge this is the first study report of the recombinant tE2 of BVDV expressed in D. melanogaster system as an antigen for ELISA.
已开发出一种简单可靠的间接酶联免疫吸附测定法，用于检测针对主要牛病毒性腹泻病毒（BVDV）免疫原E2糖蛋白的抗体（tE2-ELISA），该方法使用在果蝇系统中表达的重组C端截短E2糖蛋白（tE2）。该策略表明，tE2能有效地分泌到上清液中，无需纯化步骤，易于生产，并能进行保持其天然构象所需的翻译后修饰。使用tE2-ELISA对183份牛血清样本进行的初步分析显示，与标准同源BVDV病毒中和试验相比，其特异性为98%，敏感性为100%。结果还表明，tE2具有免疫反应性，因为原始E2的构象和抗原性在很大程度上得以保留。据我们所知，这是首次关于在果蝇系统中表达的BVDV重组tE2作为ELISA抗原的研究报告。
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