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Focusing on the research and development and application of isothermal nucleic acid amplification technology
基于荧光重组酶介导等温扩增...什曼原虫核酸检测方法的建立_林红
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A method for detecting Leishmania nucleic acids based on fluorescence recombinase-aided isothermal amplification (RAA) technology has been 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 Leishmania nucleic acid detection was developed. The assay was evaluated for its detection sensitivity using recombinant plasmids with varying copy numbers of the Leishmania ITS1 gene sequence, as well as genomic DNA samples of Leishmania at different concentrations. Additionally, the assay’s specificity was assessed by performing RAA amplification reactions with genomic DNA templates derived from other blood-transmitted parasites, including Babesia rodhaini, Toxoplasma gondii, Plasmodium vivax, Plasmodium ovale, Plasmodium falciparum, Plasmodium malariae, Leishmania donovani, and Leishmania infantum.
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