Innovative Technology

Innovation

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

新型冠状病毒核酸荧光型RT-RAA检测方法的建立及其评价

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Abstract: Objective To establish a fluorescent RTRAA-based rapid detection method for the novel coronavirus (SARS-CoV-2). Methods An in vitro-transcribed RNA template of SARS-CoV-2 was used, and single-stranded DNA-binding protein, recombinase, and DNA polymerase were employed to rapidly amplify specific gene fragments—namely, the ORF7ab gene and the S gene sequence—at a constant temperature of 40.5°C. The method was preliminarily evaluated using nasopharyngeal swab samples from confirmed COVID-19-positive patients as well as from individuals with other respiratory viral infections. Results The developed method achieved detection times of less than 20 minutes for both target genes of SARS-CoV-2, with a sensitivity of 2 copies per reaction tube for each gene. Additionally, the assay demonstrated 100% specificity. The lowest detection limits for both primer sets were consistent across three independent replicate experiments, yielding identical amplification curves with nearly identical shapes. Conclusion The proposed method exhibits high sensitivity, strong specificity, excellent reproducibility, and high concordance with clinical sample results. Importantly, it does not require expensive equipment, making it well-suited for rapid, on-site testing applications.

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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

中东呼吸综合征冠状病毒RT_RAA快速检测方法的建立及应用-宁波国旅&宁波检验检疫

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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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中东呼吸综合征冠状病毒RT_RAA快速检测方法的建立及应用-宁波国旅&宁波检验检疫