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Focusing on the research and development and application of isothermal nucleic acid amplification technology
Development of a Nucleic Acid Detection Method for Leishmania spp. Based on Fluorescent Recombinase-Mediated Isothermal Amplification—Hong Lin
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A method for detecting Leishmania nucleic acids based on recombinase-mediated isothermal amplification (RAA) technology with fluorescence detection was established. Methods: Specific primers and probes targeting the internal transcribed spacer 1 (ITS1) gene of Leishmania were designed for RAA detection. Through primer pair screening and optimization of primer and probe concentrations, a fluorescent RAA assay for detecting Leishmania nucleic acids was developed. RAA amplification was performed using recombinant plasmids containing different copy numbers of the Leishmania ITS1 gene sequence and genomic DNA from Leishmania at varying concentrations as templates, to evaluate the assay’s detection sensitivity. Additionally, RAA amplification was carried out using genomic DNA from other blood-transmitted parasites, including Babesia rodentium, Toxoplasma gondii, Plasmodium vivax, Plasmodium ovale, Plasmodium falciparum, Plasmodium malariae, Leishmania donovani, and Leishmania infantum, to assess the assay’s detection specificity.
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