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Focusing on the research and development and application of isothermal nucleic acid amplification technology
Study on a Rapid Nucleic Acid Amplification Detection Method Mediated by Reverse Transcriptase Recombinase for Influenza A Virus—Zhejiang National Travel & Zhejiang Inspection and Quarantine
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Abstract: Objective To develop a rapid method for detecting influenza A virus using reverse transcription recombinase-mediated amplification (RT-RAA) technology. Methods Reverse transcriptase was used to convert influenza A virus RNA into cDNA. Primers and probes were designed based on conserved sequences of influenza A virus from the NCBI gene database. Using cDNA as a template, RT-RAA was employed to detect influenza A virus. A plasmid carrying the influenza A virus genome was constructed, and known samples were tested to evaluate the sensitivity and specificity of this method. Results The universal primers and probes for influenza A virus effectively amplified the viral nucleic acid without cross-reacting with other non-influenza viruses. The RT-RAA reaction was carried out at a constant temperature of 39°C and took only 30 minutes to yield amplification results. The lowest detectable copy number in the reaction system was 100 copies. Conclusion The established RT-RAA method for detecting influenza A virus exhibits high sensitivity and specificity, offers rapid reaction times, and is simple to perform, making it suitable for the rapid detection of influenza A virus in a universal format.
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