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Recombinase-mediated amplification rapid detection method for Tahyna virus—CDC & Yunnan Institute for Parasite Control
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Abstract: Objective: To develop a detection method for Tahyna virus (TAHV) using reverse transcription recombinase-mediated amplification (RT-RAA). Methods: Primers and probes were designed based on the conserved regions of TAHV segments. The sensitivity of the RT-RAA assay was evaluated using a plasmid containing the target gene fragment and viral cell cultures. The specificity of the assay was validated by testing seven viruses belonging to the genera Flavivirus, Alphavirus, Nairovirus, and Southeast Asian Twelve-Segment Virus. Additionally, 30 batches of mosquito and insect samples collected in 2018 from the Ningxia Hui Autonomous Region were used to evaluate the performance of this detection method. Results: The detection limit of the assay for the constructed plasmid standard was 100 copies per reaction, while the lowest detectable titer in viral cell cultures was 10 plaque-forming units per reaction. No cross-reactivity was observed with the other seven arboviruses tested. When applied to mosquito and insect samples, the RT-RAA assay showed 100% concordance with results obtained by real-time quantitative PCR. Conclusion: A rapid, specific, and sensitive RT-RAA assay for TAHV detection has been established and is suitable for on-site detection of TAHV nucleic acids in clinical specimens.
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