A two-tube multiplex real-time RT-PCR assay for the detection of four hemorrhagic fever viruses: severe fever with thrombocytopenia syndrome virus, Hantaan virus, Seoul virus, and dengue virus

被引:0
|
作者
Zhifeng Li
Xian Qi
Minghao Zhou
Changjun Bao
Jianli Hu
Bin Wu
Shenjiao Wang
Zhongmin Tan
Jianguang Fu
Jun Shan
Yefei Zhu
Fenyang Tang
机构
[1] Jiangsu Provincial Center for Disease Control and Prevention,Department of Acute Infectious Diseases Control and Prevention
来源
Archives of Virology | 2013年 / 158卷
关键词
Dengue Virus; Japanese Encephalitis Virus; Viral Hemorrhagic Fever; Hemorrhagic Fever With Renal Syndrome; Japanese Encephalitis Virus;
D O I
暂无
中图分类号
学科分类号
摘要
The aim of this study was to develop and evaluate a two-tube multiplex real-time RT-PCR assay for the detection and identification of four viral hemorrhagic fever (VHF) pathogens, severe fever with thrombocytopenia syndrome virus (SFTSV), Hantaan virus (HTNV), Seoul virus (SEOV), and dengue virus (DENV), from human clinical samples. The two-tube multiplex real-time RT-PCR assay we developed has a sensitivity of 10 copies/μL for each of the targets, and the performance was linear within the range of at least 107 transcript copies. Moreover, we evaluated the specificity of the assay using other virus RNA as template, and found no cross-reactivity. This new assay is able to detect SFTSV, HTNV, SEOV and DENV in two reactions and brings a cost of 40 % compared to separate reactions. Evaluation of this assay with clinical serum samples from laboratory-confirmed patients and healthy donors showed 100 % clinical diagnostic sensitivity and over 99 % specificity. The assay was applied for scanning 346 clinical samples collected from patients admitted to the hospital with suspected VHF and compared with virus isolation and immunofluorescence assay (IFA). The assay indentified 59 SFTSV-, 12 HTNV-, 11 SEOV- and 9 DENV-positive samples and showed higher sensitivity. This assay thus provides a reliable and cost-effective screening tool for early clinical diagnosis of SFTSV, HTNV, SEOV and DENV in the acute phase.
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页码:1857 / 1863
页数:6
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