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一种用于快速定量甲型/ H3N2流感神经氨酸酶蛋白的灵敏且特异的ELISA免疫捕获检测法。

A sensitive and specific ELISA immunocapture assay for rapid quantitation of influenza A/H3N2 neuraminidase protein.

作者信息

Gerentes L, Kessler N, Aymard M

机构信息

W.H.O., National Influenza Centre, Laboratoire de Virologie, Lyon, France.

出版信息

J Virol Methods. 1998 Aug;73(2):185-95. doi: 10.1016/s0166-0934(98)00056-1.

Abstract

Both HA and NA proteins elicit antibodies which have been shown to be capable of altering the course of infection. Nevertheless, while influenza virus vaccine standardization involves hemagglutinin (HA) and neuraminidase (NA) in terms of antigenic characterization, only HA protein quantitation is undertaken. An immunocapture ELISA (EIA) is described for N2 NA quantitation, based on the use of a highly specific monoclonal antibody (MAb) for capturing NA and an anti-NA antiserum for antigen detection. The amounts of NA in samples were deduced from the standard curve established by using purified NA. The NA-EIA is specific and detects as a little as 7 ng/ml. The capture and detector antibodies directed against A/Beijing/32/92 NA were shown to react with H3N2 prototype strains used in current influenza vaccines, provided that an antigenically matched reference NA is used as standard.

摘要

血凝素(HA)和神经氨酸酶(NA)蛋白均可诱导产生抗体,这些抗体已被证明能够改变感染进程。然而,尽管流感病毒疫苗标准化在抗原特性方面涉及血凝素(HA)和神经氨酸酶(NA),但仅对HA蛋白进行定量。本文描述了一种免疫捕获酶联免疫吸附测定法(EIA)用于N2 NA定量,该方法基于使用一种高度特异性的单克隆抗体(MAb)捕获NA以及一种抗NA抗血清进行抗原检测。样品中NA的含量通过使用纯化的NA建立的标准曲线推导得出。NA-EIA具有特异性,检测下限低至7 ng/ml。针对A/北京/32/92 NA的捕获抗体和检测抗体被证明可与当前流感疫苗中使用的H3N2原型毒株发生反应,前提是使用抗原匹配的参考NA作为标准品。

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