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基于单克隆抗体的 IgG 和 IgM ELISA 的开发用于诊断严重发热伴血小板减少综合征病毒感染。

Development of monoclonal antibody based IgG and IgM ELISA for diagnosis of severe fever with thrombocytopenia syndrome virus infection.

机构信息

Guizhou University, School of Medicine, Guiyang, China.

Center for Disease Control and Prevention of Shanxi Province, Xian, China.

出版信息

Braz J Infect Dis. 2022 Jul-Aug;26(4):102386. doi: 10.1016/j.bjid.2022.102386. Epub 2022 Jul 11.

DOI:10.1016/j.bjid.2022.102386
PMID:35835158
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC9459026/
Abstract

INTRODUCTION

Severe fever with thrombocytopenia syndrome virus (SFTSV) is a newly emerged virus that poses a great threat to human health because of high fatality rate.

METHODS

To develop sensitive and specific sero-diagnostic systems for SFTSV infections, monoclonal antibodies (MAbs) against recombinant SFTSV nucleocapsid (rSFTSV-N) protein were developed by immunizing BALB/C mice with rSFTSV-N protein and fusing the spleen cells with SP2/0 myeloma cells. Three hybridoma cell lines secreting MAbs against rSFTSV-N were obtained. MAb based IgG sandwich enzyme linked immunosorbent assay (ELISA) and IgM capture ELISA systems were established by using the newly developed MAbs. One hundred fifteen clinical suspected SFTS patients serum samples were used to evaluate the newly established systems by comparing with the total antibody detecting sandwich ELISA system and indirect ELISA systems.

RESULTS

The MAbs based sandwich IgG ELISA was perfectly matched with that of the total antibody sandwich ELISA and the indirect IgG ELISA. IgM capture ELISA results perfectly matched with that of the total antibody sandwich ELISA while detecting eight additional positive samples missed by the indirect IgM ELISA.

CONCLUSIONS

The MAbs against rSFTSV-N protein offer new tools for SFTSV studies and our newly developed MAb-based IgG and IgM capture ELISA systems would offer safe and useful tools for diagnosis of SFTS virus infections and epidemiological investigations.

摘要

简介

严重发热伴血小板减少综合征病毒（SFTSV）是一种新出现的病毒，由于其高死亡率，对人类健康构成了巨大威胁。

方法

为了开发针对 SFTSV 感染的敏感和特异的血清学诊断系统，我们用 rSFTSV-N 蛋白免疫 BALB/C 小鼠，并将其脾细胞与 SP2/0 骨髓瘤细胞融合，制备了针对重组 SFTSV 核衣壳（rSFTSV-N）蛋白的单克隆抗体（MAbs）。获得了 3 株分泌针对 rSFTSV-N 的 MAbs 的杂交瘤细胞系。用新开发的 MAb 建立了基于 MAb 的 IgG 夹心酶联免疫吸附试验（ELISA）和 IgM 捕获 ELISA 系统。通过与总抗体检测夹心 ELISA 系统和间接 ELISA 系统进行比较，用 115 份临床疑似 SFTS 患者血清样本评估新建立的系统。

结果

基于 MAb 的夹心 IgG ELISA 与总抗体夹心 ELISA 和间接 IgG ELISA 完全匹配。IgM 捕获 ELISA 的结果与总抗体夹心 ELISA 完全匹配，同时检测到间接 IgM ELISA 漏掉的 8 个额外阳性样本。

结论

针对 rSFTSV-N 蛋白的 MAb 为 SFTSV 研究提供了新的工具，我们新开发的基于 MAb 的 IgG 和 IgM 捕获 ELISA 系统将为 SFTS 病毒感染的诊断和流行病学调查提供安全有用的工具。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ecc0/9459026/a0e3ee2d49ee/gr2.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ecc0/9459026/53bd4fe6324d/gr1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ecc0/9459026/a0e3ee2d49ee/gr2.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ecc0/9459026/53bd4fe6324d/gr1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ecc0/9459026/a0e3ee2d49ee/gr2.jpg

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