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Focusing on the research and development and application of isothermal nucleic acid amplification technology
Reverse Transcription Recombinase-Aided Amplification Assay for West Nile Virus—Zhejiang National Travel & Beijing National Travel
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Abstract: [Background] With the widespread global prevalence of West Nile fever, the risk of its introduction into China has increased. Developing a rapid detection method for West Nile virus will help establish a methodological reserve for its detection. [Objective] To establish a reverse transcription recombinase-aided amplification (RT-RAA) assay for the detection of West Nile virus. [Methods] Primers and probes were designed based on conserved regions of the West Nile virus genome to develop an RT-RAA assay for West Nile virus detection. The repeatability, specificity, and sensitivity of this method were evaluated. [Results] The entire amplification reaction process of the RT-RAA method is conducted at a constant temperature of 39°C, with a short detection time (within 20 minutes). The detection limit can reach as low as 10 copies. The assay shows no cross-reactivity with mosquito-borne viruses such as Chikungunya virus, Dengue virus, Japanese encephalitis virus, and Yellow fever virus, demonstrating excellent specificity. Sample testing results were consistent with expectations. [Conclusion] The established RT-RAA assay for West Nile virus exhibits rapidity, specificity, and high sensitivity, making it suitable for rapid point-of-entry screening and epidemiological surveillance of West Nile virus.
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