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Measuring Dengue Virus RNA in the Culture Supernatant of Infected Cells by Real-time Quantitative Polymerase Chain Reaction
Published on: November 1, 2018
Development and validation of a real-time reverse transcriptase PCR assay for sensitive detection of SFTSV
Peibin Zeng1, Zhendong Yang2, Sonia Bakkour3
1West China School of Public Health, Sichuan University, Chengdu, Sichuan, China.
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.
