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Development of a Nucleic Acid Detection Method for Echinococcus granulosus Based on Recombinase-Aided Isothermal Amplification Technology—Jiangsu Institute of Blood-Related Diseases
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A nucleic acid detection method for Echinococcus granulosus based on recombinase-mediated isothermal amplification (RAA) technology was established. Methods: Specific RAA amplification primers and a fluorescent detection probe targeting a 12S rRNA gene fragment of E. granulosus were designed, screened, and synthesized to develop a fluorescent RAA assay for E. granulosus detection. Fluorescent RAA amplification was performed using recombinant plasmids containing different copy numbers of the target sequence and genomic DNA from E. granulosus at varying concentrations as templates, and the detection sensitivity was evaluated. Additionally, fluorescent RAA amplification was conducted using genomic DNA from E. granulosus, Multiceps multilocularis, Schistosoma japonicum, Schistosoma mansoni, Ancylostoma duodenale, Clonorchis sinensis, Taenia saginata, Diphyllobothrium latum, and Taenia solium as templates, and the assay’s specificity was assessed.
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