Serological diagnosis of hantavirus infections by an enzyme-linked immunosorbent assay based on detection of immunoglobulin G and M responses to recombinant nucleocapsid proteins of five viral serotypes
Serological diagnosis of hantavirus infections by an enzyme-linked immunosorbent assay based on detection of immunoglobulin G and M responses to recombinant nucleocapsid proteins of five viral serotypes
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DOI：
10.1128/jcm.35.5.1122-1130.1997
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发表时间：
1997-05-01
影响因子：
9.4
通讯作者：
Juto, P
中科院分区：
文献类型：
--
作者：
Elgh, F;Lundkvist, A;Juto, P
在世界范围内，汉他病毒每年导致超过10万人感染。快速和准确的方法对监测急性感染和流行病学研究都很重要。我们和其他人已经表明，汉坦病毒核衣壳蛋白（Ns）的氨基末端是检测汉坦病毒疾病中特异性抗体的敏感工具。因此，我们在大肠杆菌中表达了已知对人致病的五种汉坦病毒的截短的N（氨基酸1至117）;汉坦（HTN）、首尔（SEO）、多布拉瓦（DOE）、辛农布尔（SN）和普马拉（PUU）病毒。为了获得用于酶联免疫吸附测定（ELISA）的纯抗原，通过多组氨酸-金属螯合物亲和层析纯化重组蛋白。来自斯堪的纳维亚、斯洛文尼亚、俄罗斯、韩国、中国和美国的汉他病毒感染者的多克隆动物抗血清和一组血清标本被用于评价该方法的有效性。对于人和动物血清，可以将抗体应答分为两组：一方面具有HTN、SEO和DOE病毒反应性，另一方面具有SN和PUU病毒反应性。在斯堪的纳维亚、俄罗斯欧洲部分和美国的血清中，抗体应答主要针对PUU和SN病毒群。来自亚洲的血清几乎只与HTN、SEO和DOB型病毒反应。这是真实的免疫球蛋白M（IgM）和IgG抗体反应，表明这种类型的歧视，可以在汉坦病毒感染的急性期。在来自巴尔干半岛的地区（斯洛文尼亚）的患者中发现HTN、SEO和DOE病毒以及PUU和SN病毒类型的抗体应答模式。
Worldwide, hantaviruses cause more than 100,000 human infections annually. Rapid and accurate methods are important both in monitoring acute infections and for epidemiological studies. We and others have shown that the amino termini of hantavirus nucleocapsid proteins (Ns) are sensitive tools for the detection of specific antibodies in hantavirus disease. Accordingly, we expressed truncated Ns (amino acids 1 to 117) in Escherichia coli from the five hantaviruses known to be pathogenic to man; Hantaan (HTN), Seoul (SEO), Dobrava (DOE), Sin Nombre (SN), and Puumala (PUU) viruses. In order to obtain pure antigens for use in an enzyme-linked immunosorbent assay (ELISA), the recombinant proteins were purified by polyhistidine-metal chelate affinity chromatography. Polyclonal animal antisera and a panel of serum specimens from hantavirus-infected individuals from Scandinavia, Slovenia, Russia, Korea, China, and the United States were used to evaluate the usefulness of the method. With both human and animal sera, it was possible to designate the antibody response into two groups: those with HTN, SEO, and DOE virus reactivity on the one hand and those with SN and PUU virus reactivity on the other. In sera from Scandinavia, European Russia, and the United States, the antibody response was directed mainly to the PUU and SN virus group. The sera from Asia reacted almost exclusively with the HTN, SEO, and DOB types of viruses. This was true for both the immunoglobulin M (IgM) and IgG antibody responses, indicating that this type of discrimination can be done during the acute phase of hantavirus infections. Both the HTN, SEO, and DOE virus and the PUU and SN virus types of antibody response patterns were found in patients from the Balkan region (Slovenia).