title: Detection of severe fever with thrombocytopenia syndrome virus by reverse transcription-cross-priming amplification coupled with vertical flow visualization.
abstract: A virus known as severe fever with thrombocytopenia syndrome virus (SFTSV) was recently identified as the etiological agent of severe fever with thrombocytopenia syndrome (SFTS) in China. Reliable laboratory detection and identification of this virus are likely to become clinically and epidemiologically desirable. We developed a nearly instrument-free, simple molecular method which incorporates reverse transcription-cross-priming amplification (RT-CPA) coupled with a vertical flow (VF) visualization strip for rapid detection of SFTSV. The RT-CPA-VF assay targets a conserved region of the M segment of the SFTSV genome and has a limit of detection of 100 copies per reaction, with no cross-reaction with other vector-borne bunyaviruses and bacterial pathogens. The performance of the RT-CPA-VF assay was determined with 175 human plasma specimens collected from 89 clinically suspected SFTS patients and 86 healthy donors. The sensitivity and specificity of the assay were 94.1% and 100.0%, respectively, compared with a combination of virus culture and real-time RT-PCR. The entire procedure, from specimen processing to result reporting, can be completed within 2 h. The simplicity and nearly instrument-free platform of the RT-CPA-VF assay make it practical for point-of-care testing.
authors.affiliation: Institute of Pathogen Microbiology, Jiangsu Provincial Center for Disease Prevention and Control, Key Laboratories of Enteric Pathogenic Microbiology, Ministry of Health, Nanjing, China.
authors.forename: Lunbiao
authors.initials: L
authors.lastname: Cui
authors.forename: Yiyue
authors.initials: Y
authors.lastname: Ge
authors.forename: Xian
authors.initials: X
authors.lastname: Qi
authors.forename: Gaolian
authors.initials: G
authors.lastname: Xu
authors.forename: Haijing
authors.initials: H
authors.lastname: Li
authors.forename: Kangchen
authors.initials: K
authors.lastname: Zhao
authors.forename: Bin
authors.initials: B
authors.lastname: Wu
authors.forename: Zhiyang
authors.initials: Z
authors.lastname: Shi
authors.forename: Xiling
authors.initials: X
authors.lastname: Guo
authors.forename: Lin
authors.initials: L
authors.lastname: Hu
authors.forename: Qimin
authors.initials: Q
authors.lastname: You
authors.forename: Li-Hong
authors.initials: LH
authors.lastname: Zhang
authors.forename: Alexander N
authors.initials: AN
authors.lastname: Freiberg
authors.forename: Xuejie
authors.initials: X
authors.lastname: Yu
authors.forename: Hua
authors.initials: H
authors.lastname: Wang
authors.forename: Minghao
authors.initials: M
authors.lastname: Zhou
authors.forename: Yi-Wei
authors.initials: YW
authors.lastname: Tang
mesh_terms: D002044:Bunyaviridae Infections / Q000175:diagnosis*; D002681:China; D045142:Cross-Priming; D006801:Humans; D025202:Molecular Diagnostic Techniques / Q000379:methods*; D021141:Nucleic Acid Amplification Techniques / Q000379:methods*; D016856:Phlebovirus / Q000235:genetics / Q000302:isolation & purification*; D048348:Reverse Transcription; D012680:Sensitivity and Specificity; D014773:Virology / Q000379:methods*