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基于三种有代表性的汉坦病毒截短核衣壳抗原的免疫层析条检测方法的建立。

Development of an immunochromatography strip test based on truncated nucleocapsid antigens of three representative hantaviruses.

机构信息

Department of Microbiology, Graduate School of Medicine, Hokkaido University, Kita 15, Nishi 7, Kita ku, Sapporo 060-8683, Japan.

出版信息

Virol J. 2014 May 14;11:87. doi: 10.1186/1743-422X-11-87.

Abstract

BACKGROUND

Hantaviruses are causative agents of hemorrhagic fever with renal syndrome (HFRS) and nephropathia epidemica (NE) in the Old World and hantavirus pulmonary syndrome (HPS) in the New World. There is a need for time-saving diagnostic methods. In the present study, recombinant N antigens were used as antigens in an immunochromatography strip (ICG) test to detect specific IgG antibodies.

METHODS

The N-terminal 103 amino acids (aa) of Hantaan virus (HTNV), Puumala virus (PUUV) and Andes virus (ANDV) nucleocapsid (N) protein were expressed in E. coli as representative antigens of three groups (HFRS, NE and HPS-causing viruses) of hantavirus. Five different types of ICG test strips, one antigen line on one strip for each of the three selected hantaviruses (HTNV, PUUV and ANDV), three antigen lines on one strip and a mixed antigen line on one strip, were developed and sensitivities were compared.

RESULTS

A total of 87 convalescent-phase patient sera, including sera from 35 HFRS patients, 36 NE patients and 16 HPS patients, and 25 sera from healthy seronegative people as negative controls were used to evaluate the ICG test. Sensitivities of the three-line strip and mixed-line strip were similar to those of the single antigen strip (97.2 to 100%). On the other hand, all of the ICG test strips showed high specificities to healthy donors.

CONCLUSION

These results indicated that the ICG test with the three representative antigens is an effective serodiagnostic tool for screening and typing of hantavirus infection in humans.

摘要

背景

汉坦病毒是旧世界引起肾综合征出血热(HFRS)和流行性肾病(NE)、新世界引起汉坦病毒肺综合征(HPS)的病原体。因此需要开发省时的诊断方法。本研究中,我们使用重组 N 抗原作为免疫层析检测条(ICG)试验的抗原,以检测特异性 IgG 抗体。

方法

汉滩病毒(HTNV)、普马拉病毒(PUUV)和安第斯病毒(ANDV)核衣壳(N)蛋白的 N 末端 103 个氨基酸(aa)在大肠杆菌中表达,作为三组(HFRS、NE 和 HPS 致病病毒)汉坦病毒的代表性抗原。我们开发了 5 种不同类型的 ICG 检测条,每条检测条上有 1 个抗原线,用于检测 3 种选定的汉坦病毒(HTNV、PUUV 和 ANDV),每条检测条上有 3 个抗原线和 1 个混合抗原线,并比较了它们的敏感性。

结果

我们共使用了 87 份恢复期患者血清,包括 35 份 HFRS 患者血清、36 份 NE 患者血清和 16 份 HPS 患者血清,以及 25 份健康血清阴性对照者血清,用于评估 ICG 检测。3 线条和混合线条的敏感性与单抗原条相似(97.2%~100%)。另一方面,所有的 ICG 检测条对健康供体均显示出很高的特异性。

结论

这些结果表明,使用 3 种代表性抗原的 ICG 检测是一种有效的血清学诊断工具,可用于筛查和鉴定人类汉坦病毒感染。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0716/4047433/6459e6778963/1743-422X-11-87-1.jpg

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