Innovation
Focusing on the research and development and application of isothermal nucleic acid amplification technology
甲型流感病毒的逆转录重组酶介导核酸扩增快速检测方法研究-浙江国旅&浙江检验检疫
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Abstract: Objective To establish a rapid method for detecting influenza A virus using reverse transcription recombinase-mediated amplification (RT-RAA) technology. Methods Reverse transcriptase was employed to convert influenza A virus RNA into cDNA. Primers and probes were designed based on the conserved sequences of influenza A virus available in the NCBI gene database. The cDNA served as the template for RT-RAA detection of influenza A virus. Additionally, a plasmid carrying the influenza A virus sequence was constructed to validate the method by testing known samples, thereby evaluating its sensitivity and specificity. Results Using influenza A virus–specific primers and probes, the assay successfully amplified viral nucleic acids without cross-reacting with non-influenza viruses. The entire RT-RAA reaction process was completed at a constant temperature of 39°C within 30 minutes, yielding clear amplification results. The lowest detectable copy number in the reaction system was 100 copies. Conclusion The established RT-RAA method demonstrates high sensitivity and specificity, offers rapid results, and is simple to perform, making it well-suited for the quick detection of influenza A virus in general populations.
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