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

A Study on the Recombinase-Mediated Detection Methodology for the Avian Influenza Virus H5N1 – Chen Shudan

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A rapid method for detecting the avian influenza virus H5N1 was established using reverse transcription-recombinase-mediated amplification (RT-RAA) technology. Primers and probes were designed based on the conserved sequences of the H5N1 avian influenza virus. The H5N1 RNA was first reverse-transcribed into cDNA, which then served as a template for amplifying and detecting the viral nucleic acid sequence. In this study, a plasmid containing the H5N1 nucleic acid sequence was constructed and used as a template to detect plasmids at various concentrations, thereby analyzing the sensitivity of the method. The specificity of the method was verified by testing known samples. The H5N1-specific primers and probes developed in this study can efficiently amplify the corresponding viral nucleic acid within 10 to 30 minutes at a constant temperature of 39°C, with no cross-reactivity to other influenza viruses. The detection sensitivity for the plasmid reached as low as 10 copies. The established RT-RAA method for detecting the H5N1 avian influenza virus exhibits both high sensitivity and high specificity. Moreover, the method is simple to perform and is well-suited for the rapid detection of the H5N1 avian influenza virus.