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Development of a Nucleic Acid Detection Method for Angiostrongylus cantonensis Based on Recombinase-Aided Isothermal Amplification Technology—Jiangsu Institute of Parasitic Diseases
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A nucleic acid detection method for Angiostrongylus cantonensis based on recombinase-mediated isothermal amplification (RAA) technology was established. Methods: The internal transcribed spacer 1 (ITS1) gene sequence of Angiostrongylus cantonensis was selected as the target gene sequence. Specific primers and probes with the highest specificity were designed, synthesized, and screened. A basic RAA detection method and a fluorescent RAA detection method were then developed for the detection of Angiostrongylus cantonensis. Fluorescent RAA amplification was performed using recombinant plasmids containing different copy numbers of the target gene fragment sequence and genomic DNA from Angiostrongylus cantonensis at varying concentrations as templates, and the detection sensitivity was evaluated. Additionally, fluorescent RAA amplification was carried out using genomic DNA extracted from the bodies of Angiostrongylus cantonensis, Schistosoma mansoni, Ascaris lumbricoides, Clonorchis sinensis, Echinococcus granulosus, Ancylostoma duodenale, Biomphalaria glabrata, and Bithynia fuchsi as templates, and the detection specificity was assessed.
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