Innovation
Focusing on the research and development and application of isothermal nucleic acid amplification technology
塔希纳病毒的重组酶介导扩增快速检测方法-CDC&云南寄生虫防治所
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Abstract: Objective: To establish a detection method for Tahyna virus (TAHV) using reverse transcription recombinase-aided amplification (RT-RAA). Methods: Primers and probes were designed targeting the conserved region of TAHV segments. The sensitivity of the RT-RAA assay was evaluated using plasmids containing the target gene fragment and viral cell cultures, while its specificity was validated by testing seven viruses belonging to the Flavivirus, Alphavirus, Peribunyavirus, and Southeast Asian Twelve-Segment Virus genera. Additionally, 30 batches of mosquito samples collected in 2018 from the Ningxia Hui Autonomous Region were used to assess the performance of this detection method. Results: The assay demonstrated a detection limit of 100 copies per reaction for the constructed plasmid standard and a minimum detection threshold of 10 plaque-forming units per reaction in viral cultures. No cross-reactivity was observed with the other seven arboviruses tested. When applied to mosquito samples, the RT-RAA method showed 100% agreement with results obtained by real-time quantitative PCR. Conclusion: A rapid, specific, and highly sensitive RT-RAA assay for TAHV detection has been successfully established, making it suitable for on-site nucleic acid testing of TAHV in clinical specimens.
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