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

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

Author information

1
West China School of Public Health, Sichuan University, Chengdu, Sichuan, China.
2
Xinyang 154 Military Hospital, Xinyang, China.
3
Blood Systems Research Institute, San Francisco, California.
4
Institute of Blood Transfusion, Chinese Academy of Medical Sciences, Chengdu, Sichuan, China.
5
Department of Pathology, Stanford University, Palo Alto, California.
6
Department of Pathology, Johns Hopkins University, Baltimore, Maryland.

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

PMID:
28036115
PMCID:
PMC5422136
DOI:
10.1002/jmv.24760
[Indexed for MEDLINE]
Free PMC Article

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