Parasites, Hosts and Diseases

The Korea Society for Parasitology and Tropical Medicine (대한기생충학·열대의학회)
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A Fluorescent Recombinase Aided Amplification Assay for Detection of Babesia microti

  • Lin, Hong (Jiangsu Province Blood Center) ;
  • Zhao, Song (Key Laboratory of National Health and Family Planning Commission on Parasitic Disease Control and Prevention, Jiangsu Provincial Key Laboratory on Parasite and Vector Control Technology, Jiangsu Institute of Parasitic Diseases) ;
  • Ye, Yuying (Key Laboratory of National Health and Family Planning Commission on Parasitic Disease Control and Prevention, Jiangsu Provincial Key Laboratory on Parasite and Vector Control Technology, Jiangsu Institute of Parasitic Diseases) ;
  • Shao, Lei (Jiangsu Province Blood Center) ;
  • Jiang, Nizhen (Jiangsu Province Blood Center) ;
  • Yang, Kun (Key Laboratory of National Health and Family Planning Commission on Parasitic Disease Control and Prevention, Jiangsu Provincial Key Laboratory on Parasite and Vector Control Technology, Jiangsu Institute of Parasitic Diseases)
  • Received : 2022.03.17
  • Accepted : 2022.06.07
  • Published : 2022.06.30
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Abstract

Babesia microti is one of the most common causative agents of babesiosis. A sensitive and rapid detection is necessary for screening potentially infected individuals. In this study, B. microti cytochrome c oxidase subunit I (cox1) was selected as the target gene, multiple primers were designed, and optimized by a recombinase-aided amplification (RAA) assay. The optimal primers and probe were labeled with fluorescein. The sensitivity of fluorescent RAA (fRAA) was evaluated using gradient diluents of the cox1 recombinant plasmid and genomic DNA extracted from whole blood of B. microti infected mice. The specificity of fRAA was assessed by other transfusion transmitted parasites. The analytical sensitivity of the fRAA assay was 10 copies of recombinant plasmid per reaction and 10 fg/µl B. microti genomic DNA. No cross-reaction with any other blood-transmitted parasites was observed. Our results demonstrated that the fRAA assay would be rapid, sensitive, and specific for the detection of B. microti.

Keywords

Acknowledgement

This work was supported by Social Development Project (BE2019755) and International Technology Corporation Project (BZ2020003) from the Department of Science and Technology of Jiangsu Province. Supplementary material associated with this article can be found in the online version.

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