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Focusing on the research and development and application of isothermal nucleic acid amplification technology

禽流感病毒H5N1的重组酶介导检测方法学研究-陈淑丹

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A rapid method for detecting avian influenza virus H5N1 has been established using Reverse Transcription Recombinase-Aided Amplification (RT-RAA) technology. Primers and probes were designed based on the conserved sequences of the H5N1 avian influenza virus, enabling the reverse transcription of H5N1 RNA into cDNA. The cDNA was then used as a template to amplify and detect the specific viral nucleic acid sequence. In this study, a plasmid containing the H5N1 nucleic acid sequence was constructed as a reference standard to assess the method's sensitivity across various plasmid concentrations. Additionally, the specificity of the assay was validated by testing it against known samples. The H5N1-specific primers and probe designed in this study allow efficient amplification of the target viral nucleic acid within 10 to 30 minutes at a constant temperature of 39°C, with no cross-reactivity observed against other influenza viruses. Furthermore, the detection sensitivity for the plasmid template reached as low as 10 copies. Overall, the developed RT-RAA method demonstrates high sensitivity and specificity, while its straightforward operation makes it well-suited for the rapid detection of avian influenza virus H5N1.

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