A novel mu-capture enzyme-linked immunosorbent assay based on recombinant proteins for sensitive and specific diagnosis of hemorrhagic fever with renal syndrome
A novel mu-capture enzyme-linked immunosorbent assay based on recombinant proteins for sensitive and specific diagnosis of hemorrhagic fever with renal syndrome
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一种基于重组蛋白的新型 mu-capture 酶联免疫吸附测定法，用于灵敏、特异地诊断肾综合征出血热
DOI：
10.1128/jcm.31.5.1194-1199.1993
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发表时间：
1993
影响因子：
9.4
通讯作者：
G. Darai
中科院分区：
文献类型：
--
作者：
L. Zöller;Si;P. Gött;E. Bautz;G. Darai
汉坦病毒核衣壳蛋白最近被确定为一种主要抗原，可在肾综合征出血热患者中诱导早期且持久的体液免疫反应。开发了一种 mu 捕获酶联免疫吸附测定法，利用汉坦病毒株 Hantaan 76-118（Hantaan 血清型）和 CG 18-20（Puumala 血清型）的重组核衣壳蛋白作为诊断抗原，并使用特异性单克隆抗体作为检测系统。带组氨酸尾的重组蛋白在大肠杆菌中表达，并通过镍螯合树脂上的亲和层析一步纯化。该测定采用来自不同地理区域的肾综合征出血热患者的一组血清进行了评估。 mu-capture酶联免疫吸附测定（均为重组抗原）的总体灵敏度为100%，其特异性也为100%。免疫球蛋白 M 抗体最早在发病第 3 天就被检测到，并在发病后第 8 天至 25 天之间获得最大滴度。检测结果通常在疾病急性期后 3 至 4 个月内呈阳性，但在某些情况下长达 2 年。
Hantavirus nucleocapsid protein has recently been identified as a major antigen inducing an early and long-lasting humoral immune response in patients with hemorrhagic fever with renal syndrome. A mu-capture enzyme-linked immunosorbent assay utilizing recombinant nucleocapsid proteins of Hantavirus strains Hantaan 76-118 (Hantaan serotype) and CG 18-20 (Puumala serotype) as diagnostic antigens and specific monoclonal antibodies as the detection system has been developed. Histidine-tailed recombinant proteins were expressed in Escherichia coli and purified in a single step by affinity chromatography on a nickel-chelate resin. The assay was evaluated with a panel of sera from patients with hemorrhagic fever with renal syndrome originating from various geographic regions. The overall sensitivity of the mu-capture enzyme-linked immunosorbent assay (both recombinant antigens) was 100%, and its specificity was also found to be 100%. Immunoglobulin M antibodies were detected as early as on day 3, and maximum titers were obtained between days 8 and 25 after onset of the disease. The assay was regularly found to be positive within 3 to 4 months but in some cases up to 2 years after the acute phase of the disease.