Development and validation of a real-time reverse transcriptase PCR assay for sensitive detection of SFTSV

Peibin Zeng, Zhendong Yang, Sonia Bakkour, Bingjun Wang, Shuli Qing, Jingxing Wang, Limin Chen, Michael Busch, Hua Shan, Jing Liu, Tzong Hae Lee

Research output: Contribution to journalArticle

Abstract

Background: 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. Materials and methods: 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. Results: 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. Conclusion: 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.

Original languageEnglish (US)
JournalJournal of Medical Virology
DOIs
StateAccepted/In press - 2017

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Phlebovirus
Reverse Transcriptase Polymerase Chain Reaction
Nucleic Acids
Real-Time Polymerase Chain Reaction
Thrombocytopenia
Fever
RNA-Directed DNA Polymerase
Blood Donors
Donor Selection
Antibodies
RNA Viruses
Ticks
Limit of Detection
China

Keywords

  • Bunyavirus
  • RNA extraction
  • RNA purification
  • Test statistics

ASJC Scopus subject areas

  • Infectious Diseases
  • Virology

Cite this

Development and validation of a real-time reverse transcriptase PCR assay for sensitive detection of SFTSV. / Zeng, Peibin; Yang, Zhendong; Bakkour, Sonia; Wang, Bingjun; Qing, Shuli; Wang, Jingxing; Chen, Limin; Busch, Michael; Shan, Hua; Liu, Jing; Lee, Tzong Hae.

In: Journal of Medical Virology, 2017.

Research output: Contribution to journalArticle

Zeng, P, Yang, Z, Bakkour, S, Wang, B, Qing, S, Wang, J, Chen, L, Busch, M, Shan, H, Liu, J & Lee, TH 2017, 'Development and validation of a real-time reverse transcriptase PCR assay for sensitive detection of SFTSV', Journal of Medical Virology. https://doi.org/10.1002/jmv.24760
Zeng, Peibin ; Yang, Zhendong ; Bakkour, Sonia ; Wang, Bingjun ; Qing, Shuli ; Wang, Jingxing ; Chen, Limin ; Busch, Michael ; Shan, Hua ; Liu, Jing ; Lee, Tzong Hae. / Development and validation of a real-time reverse transcriptase PCR assay for sensitive detection of SFTSV. In: Journal of Medical Virology. 2017.
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AU - Qing, Shuli

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