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

J Med Virol. 2017 Jul;89(7):1131-1138. doi: 10.1002/jmv.24760. Epub 2017 Feb 27.

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.

Keywords: RNA extraction; RNA purification; bunyavirus; test statistics.

Publication types

  • Validation Study

MeSH terms

  • Antibodies, Viral / blood
  • Blood Donors*
  • Bunyaviridae Infections / diagnosis*
  • Bunyaviridae Infections / epidemiology
  • Bunyaviridae Infections / virology
  • China / epidemiology
  • Enzyme-Linked Immunosorbent Assay
  • Humans
  • Phlebovirus / genetics
  • Phlebovirus / immunology
  • Phlebovirus / isolation & purification*
  • RNA, Viral / analysis
  • Reverse Transcriptase Polymerase Chain Reaction / methods*
  • Sensitivity and Specificity

Substances

  • Antibodies, Viral
  • RNA, Viral