title: Development and validation of a real-time reverse transcriptase PCR assay for sensitive detection of SFTSV.
abstract: Severe fever with thrombocytopenia syndrome bunyavirus (sftsv) is an emerging tick-borne rna virus recently identified as the pathogen that causes severe fever with thrombocytopenia syndrome (sfts) in china. the existing commercial nucleic acid testing (comnat) assay with a relatively high claimed limit of quantitative detection (loqd) is not capable of sensitive detection and quantitation of sftsv. Thus, a new real-time reverse transcriptase (rt)-pcr assay with improved sensitivity is needed for clinical diagnosis; it could also be used to screen blood donors if necessary.
  
  
  
  We developed a new sftsv rt-pcr nat assay (newnat). About 129 plasma samples from 93 suspected sfts patients with typical clinical symptoms were tested using an anti-sftsv total antibody elisa and both comnat and newnat. The test performance of the two nat assays was evaluated and compared.
  
  
  
  The newnat had a lower limit for quantitative testing compared to comnat. Twelve samples were comnat negative but newnat positive. Out of 35 suspected sfts patients who were comnat negative and anti-sftsv total antibody negative, four tested positive by the newnat assay and one of these four seroconverted within 2-4 days after testing newnat positive. A high correlation was observed between the cts of the newnat and comnat assays.
  
  
  
  The newnat assay was sensitive for quantitative detection of sftsv and may be applicable to clinical diagnosis and studies of the need for blood donor screening.
authors.affiliation: West China School of Public Health, Sichuan University, Chengdu, Sichuan, China.
authors.forename: Peibin
authors.identifier: 0000-0001-6668-2380
authors.initials: P
authors.lastname: Zeng
authors.affiliation: Xinyang 154 Military Hospital, Xinyang, China.
authors.forename: Zhendong
authors.initials: Z
authors.lastname: Yang
authors.affiliation: Blood Systems Research Institute, San Francisco, California.
authors.forename: Sonia
authors.initials: S
authors.lastname: Bakkour
authors.affiliation: Xinyang 154 Military Hospital, Xinyang, China.
authors.forename: Bingjun
authors.initials: B
authors.lastname: Wang
authors.affiliation: Xinyang 154 Military Hospital, Xinyang, China.
authors.forename: Shuli
authors.initials: S
authors.lastname: Qing
authors.affiliation: Institute of Blood Transfusion, Chinese Academy of Medical Sciences, Chengdu, Sichuan, China.
authors.forename: Jingxing
authors.initials: J
authors.lastname: Wang
authors.affiliation: Xinyang 154 Military Hospital, Xinyang, China.
authors.forename: Limin
authors.initials: L
authors.lastname: Chen
authors.affiliation: Blood Systems Research Institute, San Francisco, California.
authors.forename: Michael
authors.initials: M
authors.lastname: Busch
authors.affiliation: Department of Pathology, Stanford University, Palo Alto, California.
authors.forename: Hua
authors.initials: H
authors.lastname: Shan
authors.affiliation: Department of Pathology, Johns Hopkins University, Baltimore, Maryland.
authors.forename: Jing
authors.initials: J
authors.lastname: Liu
authors.affiliation: Institute of Blood Transfusion, Chinese Academy of Medical Sciences, Chengdu, Sichuan, China.
authors.forename: Tzong-Hae
authors.initials: TH
authors.lastname: Lee
mesh_terms: D000914:Antibodies, Viral / Q000097:blood; D001782:Blood Donors; D002044:Bunyaviridae Infections / Q000175:diagnosis* / Q000453:epidemiology / Q000821:virology; D002681:China / Q000453:epidemiology; D004797:Enzyme-Linked Immunosorbent Assay; D006801:Humans; D016856:Phlebovirus / Q000235:genetics / Q000276:immunology / Q000302:isolation & purification*; D012367:RNA, Viral / Q000032:analysis; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity
chemical_list: D000914:Antibodies, Viral; D012367:RNA, Viral
keywords: RNA extraction; RNA purification; bunyavirus; test statistics