A new luciferase immunoprecipitation system assay provided serological evidence for missed diagnosis of severe fever with thrombocytopenia syndrome

Shengyao Chen; Minjun Xu; Xiaoli Wu; Yuan Bai; Junming Shi; Min Zhou; Qiaoli Wu; Shuang Tang; Fei Deng; Bo Qin; Shu Shen

doi:10.1016/j.virs.2022.01.018

February 2022

Shengyao Chen, Minjun Xu, Xiaoli Wu, Yuan Bai, Junming Shi, Min Zhou, Qiaoli Wu, Shuang Tang, Fei Deng, Bo Qin and Shu Shen. A new luciferase immunoprecipitation system assay provided serological evidence for missed diagnosis of severe fever with thrombocytopenia syndrome[J]. Virologica Sinica, 2022, 37(1): 107-114. doi: 10.1016/j.virs.2022.01.018

Citation: Shengyao Chen, Minjun Xu, Xiaoli Wu, Yuan Bai, Junming Shi, Min Zhou, Qiaoli Wu, Shuang Tang, Fei Deng, Bo Qin, Shu Shen. A new luciferase immunoprecipitation system assay provided serological evidence for missed diagnosis of severe fever with thrombocytopenia syndrome .VIROLOGICA SINICA, 2022, 37(1) : 107-114. http://dx.doi.org/10.1016/j.virs.2022.01.018

一种新的荧光素酶免疫沉淀法分析方法为发热伴血小板减少综合征的漏诊提供了血清学证据

陈声耀 ^a,c ,
许敏君 ^d ,
吴晓丽 ^a ,
白源 ^a ,
史君明 ^a ,
周敏 ^a ,
吴巧丽 ^a ,
唐霜 ^a ,
邓菲 ^a,, ,
钦博 ^b,d,, ,
沈姝 ^a,,

a 中国科学院武汉病毒研究所病毒学国家重点实验室国家病毒资源库，武汉 430071，中国
b 浙江省绍兴市东街305号绍兴市妇幼医院，绍兴312000，中国
c 中国科学院大学，北京100049，中国
d 浙江大学医学院绍兴人民医院，绍兴，312000，中国

通讯作者： 邓菲, df@wh.iov.cn; 钦博, qinbo0809@hotmail.com; 沈姝, shenshu@wh.iov.cn
收稿日期： 2021-08-11
录用日期： 2021-10-12

摘要

SFTS病毒（SFTSV）感染引起的严重发热伴血小板减少综合征（SFTS）2010年在中国首次报道，致死率高达30%。通过检测病毒RNA或抗体水平来确认SFTSV感染对于SFTS的诊断和治疗至关重要。本研究建立了一种新型的荧光素酶免疫沉淀系统（LIPS），该系统pREN2质粒系统融合表达SFTSV 核蛋白NP与海参荧光素酶（Rluc），通过测定Rluc的酶活反应水平检测血液样本中抗SFTSV抗体应答水平。对2019年从浙江省绍兴市医院共采集的464份发热患者血清样本进行LIPS检测，发现4有82例病人（17.7%）为SFTSV抗体阳性。该结果也得到了免疫荧光实验验证。此外，qRT-PCR和微量中和实验显示，82例阳性病例中，有15例患者有病毒血症，10例患者血清表现病毒中和活性，1例患者血清样本中同时检测到病毒血症和中和抗体，提示至少有上述共26例患者存在对SFTSV的感染暴露。然而，回顾患者医疗诊断记录和疾病相关临床检测指标，显示这些患者均没有被诊断为SFTS，可能与他们的症状轻微或亚临床表现有关。因此，研究结果提示：很可能存在SFTS漏诊病例， SFTS的实际发病率很可能高于记录和报告的水平。研究也证明了LIPS作为一种新兴的总抗体水平检测方法，可以成为SFTSV感染病例的实验室确诊的一种有效的手段。
- 发热伴血小板减少综合征（SFTS）
- , 发热伴血小板减少综合征病毒（SFTSV）
- , 荧光素酶免疫沉淀系统（LIPS）
- , 血清学证据
- , 漏诊

A new luciferase immunoprecipitation system assay provided serological evidence for missed diagnosis of severe fever with thrombocytopenia syndrome

Shengyao Chen ^a,c ,
Minjun Xu ^d ,
Xiaoli Wu ^a ,
Yuan Bai ^a ,
Junming Shi ^a ,
Min Zhou ^a ,
Qiaoli Wu ^a ,
Shuang Tang ^a ,
Fei Deng ^a,, ,
Bo Qin ^b,d,, ,
Shu Shen ^a,,

a State Key Laboratory of Virology and National Virus Resource Center, Wuhan Institute of Virology, Chinese Academy of Sciences, Wuhan, 430071, China
b Shaoxing Women and Children's Hospital, No. 305 East Street Road, Shaoxing, 312000, China
c University of Chinese Academy of Sciences, Beijing, 100049, China
d Shaoxing People's Hospital, Zhejiang University School of Medicine, Shaoxing, 312000, China

Corresponding author: Fei Deng, df@wh.iov.cn Bo Qin, qinbo0809@hotmail.com Shu Shen, shenshu@wh.iov.cn
Received Date: 11 August 2021
Accepted Date: 12 October 2021

Abstract

Severe fever with thrombocytopenia syndrome (SFTS), caused by SFTS virus (SFTSV) infection, was first reported in 2010 in China with an initial fatality of up to 30%. The laboratory confirmation of SFTSV infection in terms of detection of viral RNA or antibody levels is critical for SFTS diagnosis and therapy. In this study, a new luciferase immunoprecipitation system (LIPS) assay based on pREN2 plasmid expressing SFTSV NP gene and tagged with Renilla luciferase (Rluc), was established and used to investigate the levels of antibody responses to SFTSV. Totally 464 serum samples from febrile patients were collected in the hospital of Shaoxing City in Zhejiang Province in 2019. The results showed that 82 of the 464 patients (17.7%) had antibody response to SFTSV, which were further supported by immunofluorescence assays (IFAs). Further, qRT-PCR and microneutralization tests showed that among the 82 positive cases, 15 patients had viremia, 10 patients had neutralizing antibody, and one had both (totally 26 patient). However, none of these patients were diagnosed as SFTS in the hospital probably because of their mild symptoms or subclinical manifestations. All the results indicated that at least the 26 patients having viremia or neutralizing antibody were the missed diagnosis of SFTS cases. The findings suggested the occurrence of SFTS and the SFTS incidence were higher than the reported level in Shaoxing in 2019, and that LIPS may provide an alternative strategy to confirm SFTSV infection in the laboratory.

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Electronic Supplementary Material

10.1016j.virs.2022.01.018-ESM.docx