Innovation
Focusing on the research and development and application of isothermal nucleic acid amplification technology
Recombinase-mediated amplification method for rapid detection of yellow fever virus—Zhejiang National Travel
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Abstract: Objective: This study employed a recombinase-mediated isothermal nucleic acid amplification method (RAA) and, using reverse transcriptase, developed a one-step isothermal nucleic acid amplification assay for yellow fever virus (RT-RAA). Methods: Primers and probes were designed based on conserved sequences in the yellow fever virus genome. The repeatability, specificity, and sensitivity of RT-RAA were established and analyzed. The developed method was used to detect yellow fever virus samples, and the results were validated by genetic sequencing. Results: In the RT-RAA amplification system for yellow fever virus, the optimal amplification effect was achieved when 40 U of reverse transcriptase was added. This method features a short detection time (<20 min) and high sensitivity, with a detection limit as low as 100 copies. It showed no cross-reactivity with mosquito-borne viruses such as dengue virus, West Nile virus, Japanese encephalitis virus, and chikungunya virus, demonstrating excellent specificity. Conclusion: The developed RT-RAA method for yellow fever virus exhibits rapid, specific, and sensitive characteristics, making it suitable for rapid point-of-entry screening of yellow fever virus.
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