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抗基孔肯雅病毒衣壳蛋白单克隆抗体用于检测近期感染的诊断潜力。

Diagnostic potential of monoclonal antibodies against the capsid protein of chikungunya virus for detection of recent infection.

作者信息

Damle R G, Jayaram N, Kulkarni S M, Nigade K, Khutwad K, Gosavi S, Parashar D

机构信息

National Institute of Virology (NIV), Microbial Containment Complex (MCC), 130/1, Sus Road, Pashan, Pune, 411021, India.

出版信息

Arch Virol. 2016 Jun;161(6):1611-22. doi: 10.1007/s00705-016-2829-4. Epub 2016 Mar 26.

Abstract

Chikungunya fever is self-limiting. However, neurological and hemorrhagic complications have been seen in recent outbreaks. The clinical manifestations of this disease are similar to those of dengue virus infection, indicating the need for differential diagnosis in areas such as India, which are endemic for both viruses. The aim of the present study was to develop monoclonal antibodies (MAbs) against Chikungunya virus (CHIKV) and assess their use in MAb-based IgM capture ELISA (MAC ELISA). The ELISA detects CHIKV-specific IgM antibodies, a marker of recent infection, in a patient's serum. One IgG1 and two IgM isotype hybrids were obtained. All of the subclones derived from the IgG1 hybrid recognized the C protein of CHIKV. The anti-C MAb ClVE4/D9 was the most promising as a detector antibody in MAC ELISA (C-MAb ELISA) yielding higher positive-to-negative (P/N) ratios. When compared with the CHIKV MAC ELISA kit developed by the National Institute of Virology (NIV), Pune (NIV MAC ELISA), the sensitivity of the test was 87.01 % with 100 % specificity. The positive and negative predictive values (PPV and NPV) were 100 % and 94.47 %, respectively. In precision testing, standard deviation (SD) and coefficient of variation (% CV) values of the C-MAb ELISA were within acceptable limits. The C-MAb ELISA detected anti-CHIKV IgM in serum of patients up to five months after the onset of infection, indicating that anti-C MAbs have strong potential for use in MAC ELISA to detect recent CHIKV infection.

摘要

基孔肯雅热是自限性疾病。然而,在近期的疫情中出现了神经和出血性并发症。该疾病的临床表现与登革病毒感染相似,这表明在印度等两种病毒都流行的地区需要进行鉴别诊断。本研究的目的是开发针对基孔肯雅病毒(CHIKV)的单克隆抗体(MAb),并评估其在基于MAb的IgM捕获ELISA(MAC ELISA)中的应用。该ELISA可检测患者血清中CHIKV特异性IgM抗体,这是近期感染的标志物。获得了一株IgG1和两株IgM同种型杂交体。源自IgG1杂交体的所有亚克隆均识别CHIKV的C蛋白。抗C MAb ClVE4/D9作为MAC ELISA(C-MAb ELISA)中的检测抗体最具前景,产生的阳性/阴性(P/N)比值更高。与浦那国家病毒研究所(NIV)开发的CHIKV MAC ELISA试剂盒(NIV MAC ELISA)相比,该检测的灵敏度为87.01%,特异性为100%。阳性和阴性预测值(PPV和NPV)分别为100%和94.47%。在精密度测试中,C-MAb ELISA的标准差(SD)和变异系数(%CV)值在可接受范围内。C-MAb ELISA可在感染发作后长达五个月的患者血清中检测到抗CHIKV IgM,这表明抗C MAb在MAC ELISA中检测近期CHIKV感染具有很强的应用潜力。

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