Technical Centre of Animal and Plant Inspection and Quarantine, Shenzhen Entry-Exit Inspection and Quarantine Bureau, Shengzhen 518001, China.
J Virol Methods. 2010 Jan;163(1):68-73. doi: 10.1016/j.jviromet.2009.08.015. Epub 2009 Sep 1.
In this study, an immunochromatographic strip (ICS) was developed for the detection of bluetongue virus (BTV) serum antibodies. Colloidal gold particles labeled with streptococcal protein G (SPG), which can bind to the F(C) fragment of mammalian immunoglobulins, were used as the detector reagent. A recombinant VP7 BTV protein and a purified rabbit anti-SPG antibody were immobilized on test and control regions of a nitrocellulose membrane, respectively. In order to evaluate the ICS, 37 sera from animals exposed to different BTV serotypes were used as positive controls. In addition, 50 positive sera against viruses other than BTV, and eight sera taken from naive healthy sheep were used to determine the specificity of the ICS. Three hundred and three field sera taken from sheep and cattle were used after the above sera had been used for validation. Compared with the competitive ELISA (c-ELISA), the specificity and sensitivity of the ICS was 97.6% and 100%, respectively. There was excellent agreement between the results obtained by c-ELISA and the ICS (kappa=0.930). As it is rapid and easy to use, the test is suitable for the serological surveillance of BTV infection in the field.
本研究旨在开发一种用于检测蓝舌病毒（BTV）血清抗体的免疫层析条（ICS）。胶体金颗粒标记有链球菌蛋白 G（SPG），可与哺乳动物免疫球蛋白的 F(C)片段结合，用作检测试剂。重组 VP7 BTV 蛋白和纯化的兔抗 SPG 抗体分别固定在硝酸纤维素膜的测试区和对照区。为了评估 ICS，使用来自接触不同 BTV 血清型的动物的 37 份血清作为阳性对照。此外，使用 50 份针对除 BTV 以外的病毒的阳性血清和 8 份来自无经验健康绵羊的血清来确定 ICS 的特异性。在使用上述血清进行验证后，使用 330 份来自绵羊和牛的现场血清。与竞争 ELISA（c-ELISA）相比，ICS 的特异性和灵敏度分别为 97.6%和 100%。c-ELISA 和 ICS 之间的结果具有极好的一致性（kappa=0.930）。由于其快速易用，该检测方法适合用于现场 BTV 感染的血清学监测。
