Use of the recombinant nonstructural 3A, 3B, and 3AB proteins of foot-and-mouth disease virus in indirect ELISA for differentiation of vaccinated and infected cattle

被引：10

作者：

Yakovleva A.S. ^{[1
]}

Shcherbakov A.V. ^{[1
]}

Kan'shina A.V. ^{[1
]}

Mudrak N.S. ^{[1
]}

Fomina T.A. ^{[1
]}

机构：

[1] Federal Center for Animal Health, Ministry of Agriculture of the Russian Federation, Vladimir

来源：

Molecular Biology | 2006年 / 40卷 / 1期

关键词：

Diagnostic sensitivity and specificity; Expression in Escherichia coli; Foot-and-mouth disease virus; Indirect ELISA; Metal-chelate affinity chromatography; Nonstructural proteins;

D O I：

10.1134/S0026893306010195

中图分类号：

学科分类号：

摘要：

Recombinant foot-and-mouth disease virus (FMDV) proteins 3A, 3B, and 3AB were produced by expressing the corresponding genes in Escherichia coli and purified by metal-chelate affinity chromatography. The recombinant proteins were used as antigens in indirect enzyme-linked immunosorbent assay (ELISA) to differentiate between vaccinated and FMD-infected animals. The following parameters were determined: working concentrations of antigens and peroxidase conjugate of cattle anti-IgG, the optimum composition of blocking buffer, and the positive-negative threshold of the reaction. Tests performed with approximately 200 serum samples taken from animals of different immunity states showed that the protocol with protein 3A as the antigen (3A-ELISA) provided the most reliable differentiation. All the newly developed systems proved to outperform the commercial Chekit FMD-3ABC kit in sensitivity, and 3A-ELISA was no less specific. © Pleiades Publishing, Inc., 2006.

引用

页码：146 / 151

页数：5

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