Innovation
Focusing on the research and development and application of isothermal nucleic acid amplification technology
重组酶介导扩增方法快速检测黄热病毒-浙江国旅
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Abstract: Objective This study aimed to develop a one-step isothermal nucleic acid amplification method (RT-RAA) for yellow fever virus detection, utilizing recombinase-mediated isothermal amplification (RAA) combined with reverse transcription. Methods Primers and probes were designed based on conserved sequences within the yellow fever virus genome, enabling the establishment and optimization of RT-RAA. The method’s repeatability, specificity, and sensitivity were evaluated, and the newly developed RT-RAA assay was subsequently applied to analyze yellow fever virus samples, with results further validated by genetic sequencing. Results Optimal amplification performance was achieved when 40 U of reverse transcriptase enzyme was added to the reaction mixture. The RT-RAA method offers rapid detection time (<20 min) and high sensitivity, with a detection limit as low as 100 copies. Importantly, this method exhibited no cross-reactivity with other mosquito-borne viruses, including dengue, West Nile, Japanese encephalitis B, and chikungunya viruses, demonstrating excellent specificity. Conclusion The established RT-RAA method for yellow fever virus detection is characterized by its speed, specificity, and high sensitivity, making it well-suited for rapid screening at border checkpoints.
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