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重组核蛋白在酶联免疫吸附测定中用于检测甲型或乙型流感病毒感染患者体内病毒特异性免疫球蛋白A(IgA)和免疫球蛋白G(IgG)抗体的应用。

Use of recombinant nucleoproteins in enzyme-linked immunosorbent assays for detection of virus-specific immunoglobulin A (IgA) and IgG antibodies in influenza virus A- or B-infected patients.

作者信息

Voeten J T, Groen J, van Alphen D, Claas E C, de Groot R, Osterhaus A D, Rimmelzwaan G F

机构信息

WHO National Influenza Centre and Institute of Virology, Erasmus University Rotterdam, 3000 DR Rotterdam, The Netherlands.

出版信息

J Clin Microbiol. 1998 Dec;36(12):3527-31. doi: 10.1128/JCM.36.12.3527-3531.1998.

Abstract

The nucleoprotein genes of influenza virus A/Netherlands/018/94 (H3N2) and influenza virus B/Harbin/7/94 were cloned into the bacterial expression vector pMalC to yield highly purified recombinant influenza virus A and B nucleoproteins. With these recombinant influenza nucleoproteins, enzyme-linked immunosorbent assays (ELISAs) were developed for the detection of influenza virus A- and B-specific immunoglobulin A (IgA) and IgG serum antibodies. Serum samples were collected at consecutive time points after the onset of clinical symptoms from patients with confirmed influenza virus A or B infections. Nucleoprotein-specific IgA antibodies were detected in 41.2% of influenza virus A-infected patients and in 66. 7% of influenza virus B-infected patients on day 6 after the onset of clinical symptoms. In serum samples taken on day 21 (influenza virus A-infected patients) or day 28 (influenza virus B-infected patients), nucleoprotein-specific IgA antibodies could be detected in 58.8 and 58.3% of influenza virus A- and B-infected patients, respectively. At the same time, IgG antibody rises were detected in 88.2% of influenza virus A-infected patients and in 95.8% of influenza virus B-infected patients. On comparison, hemagglutination inhibition assays detected antibody titer rises in 81.3 and 72.7% of patients infected with influenza viruses A and B, respectively. In contrast to the detection of nucleoprotein-specific IgG antibodies or hemagglutination-inhibiting antibodies, the detection of nucleoprotein-specific IgA antibodies does not require paired serum samples and therefore can be considered an attractive alternative for the rapid serological diagnosis of influenza.

摘要

将甲型流感病毒A/荷兰/018/94(H3N2)和乙型流感病毒B/哈尔滨/7/94的核蛋白基因克隆到细菌表达载体pMalC中,以产生高度纯化的重组甲型和乙型流感病毒核蛋白。利用这些重组流感病毒核蛋白,开发了酶联免疫吸附测定(ELISA)法,用于检测甲型和乙型流感病毒特异性免疫球蛋白A(IgA)和IgG血清抗体。在确诊感染甲型或乙型流感病毒的患者出现临床症状后的连续时间点采集血清样本。在临床症状出现后第6天,41.2%的甲型流感病毒感染患者和66.7%的乙型流感病毒感染患者检测到核蛋白特异性IgA抗体。在第21天(甲型流感病毒感染患者)或第28天(乙型流感病毒感染患者)采集的血清样本中,分别在58.8%的甲型流感病毒感染患者和58.3%的乙型流感病毒感染患者中检测到核蛋白特异性IgA抗体。同时,在88.2%的甲型流感病毒感染患者和95.8%的乙型流感病毒感染患者中检测到IgG抗体升高。相比之下,血凝抑制试验分别在81.3%的甲型流感病毒感染患者和72.7%的乙型流感病毒感染患者中检测到抗体滴度升高。与检测核蛋白特异性IgG抗体或血凝抑制抗体不同,检测核蛋白特异性IgA抗体不需要配对血清样本,因此可被视为流感快速血清学诊断的一种有吸引力的替代方法。

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