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鸭坦布苏病毒抗体免疫层析检测试纸条的研制

Development of an immunochromatographic strip for detection of antibodies against duck Tembusu virus.

机构信息

Research Center of Avian Disease, College of Veterinary Medicine of Sichuan Agricultural University, Chengdu 611130, China; Institute of Preventive Veterinary Medicine, Sichuan Agricultural University, Chengdu 611130, China.

Research Center of Avian Disease, College of Veterinary Medicine of Sichuan Agricultural University, Chengdu 611130, China; Key Laboratory of Animal Disease and Human Health of Sichuan Province, Chengdu 611130, China; Institute of Preventive Veterinary Medicine, Sichuan Agricultural University, Chengdu 611130, China.

出版信息

J Virol Methods. 2017 Nov;249:137-142. doi: 10.1016/j.jviromet.2017.08.022. Epub 2017 Aug 31.

Abstract

Duck Tembusu virus (DTMUV), a novel flavivirus, causes severe disease in ducks. There is an urgent need for a rapid and effective diagnostic method to control the spread of DTMUV. We chose the envelope (E) protein from DTMUV as an antigen and combined it with colloidal gold particles as tracers to specifically detect anti-DTMUV antibodies. Based on the double-antigen sandwich format, an immunochromatographic strip (ICS) for the rapid detection of anti-DTMUV antibodies was developed. The ICS showed a high specificity and no cross-reactivity with other sera. By detecting a serially diluted duck anti-DTMUV serum, the sensitivity of the ICS was 16-fold higher than that of the agar gel double diffusion test. Moreover, the ICS was both stable and reproducible, maintaining the same performance at 4°C for at least 6 months. To evaluate the effectiveness of the ICS, 217 duck serum samples were tested with the ICS and an indirect enzyme-linked immunosorbent assay (iELISA). The consistency ratio of positive and negative results between the two methods was 97.87% and 97.06%, respectively. The agreement between the ICS and the ELISA was 97.24%. The ICS developed in this study offers a specific, sensitive, and rapid method to detect anti-DTMUV antibodies.

摘要

鸭坦布苏病毒(DTMUV)是一种新型黄病毒,可引起鸭的严重疾病。因此,急需一种快速有效的诊断方法来控制 DTMUV 的传播。我们选择 DTMUV 的包膜(E)蛋白作为抗原,并将其与胶体金颗粒结合作为示踪物,以特异性检测抗 DTMUV 抗体。基于双抗原夹心模式,我们开发了一种用于快速检测抗 DTMUV 抗体的免疫层析试纸条(ICS)。ICS 显示出高度的特异性,与其他血清无交叉反应。通过检测一系列稀释的鸭抗 DTMUV 血清,ICS 的灵敏度比琼脂凝胶双扩散试验高 16 倍。此外,ICS 既稳定又可重复,在至少 4°C 下至少可保持 6 个月的相同性能。为了评估 ICS 的有效性,我们用 ICS 和间接酶联免疫吸附试验(iELISA)检测了 217 份鸭血清样本。两种方法的阳性和阴性结果的一致性比率分别为 97.87%和 97.06%。ICS 与 ELISA 的一致性为 97.24%。本研究开发的 ICS 提供了一种特异性、敏感性和快速的方法来检测抗 DTMUV 抗体。

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