Innovative Technology

Innovation

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

实时荧光逆转录重组酶介导的等温扩增技术检测寨卡病毒方法的建立

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[Abstract] Objective: To establish a real-time fluorescence reverse transcription-recombinase-aided amplification (RT-RAA) method for detecting Zika virus (ZIKV), enabling rapid on-site screening and diagnosis of ZIKV. Methods: Based on genomic sequence alignment, conserved ZIKV sequences were selected to design RT-RAA-specific primers and probes. The sensitivity and reproducibility of the method were validated using serially diluted ZIKV recombinant plasmids and viral RNA extracts from clinical isolates. Specificity was assessed by testing against other arboviruses, including dengue virus, chikungunya virus, and West Nile virus. Results: The established RT-RAA method allows efficient amplification of ZIKV within 30 minutes at a constant temperature of 39°C. Sensitivity analysis revealed that the assay can detect as low as 15 copies per reaction with 95% confidence. The method exhibits high specificity, showing no cross-reactivity with dengue virus, chikungunya virus, or West Nile virus, and demonstrates excellent reproducibility. Conclusion: The RT-RAA isothermal amplification method developed in this study offers several advantages, including rapid response, minimal requirement for sophisticated equipment, and straightforward operation, making it well-suited for emergency and rapid detection scenarios.

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（脑炎）-CDC.pdf

双重荧光RT-RAA检测5种蚊媒病毒方法的建立_周冬根

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双重荧光RT-RAA检测5种蚊媒病毒方法的建立_周冬根