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ANCA: artificial nucleic acid circuit with argonaute protein for one-step isothermal detection of antibiotic-resistant bacteria

Author

Listed:
  • Hyowon Jang

    (Korea Research Institute of Bioscience and Biotechnology (KRIBB))

  • Jayeon Song

    (Korea Research Institute of Bioscience and Biotechnology (KRIBB)
    Massachusetts General Hospital Research Institute
    Harvard Medical School)

  • Sunjoo Kim

    (Gyeongsang National University Hospital, Gyeongsang National University College of Medicine)

  • Jung-Hyun Byun

    (Gyeongsang National University Hospital, Gyeongsang National University College of Medicine)

  • Kyoung G. Lee

    (National NanoFab Center (NNFC))

  • Kwang-Hyun Park

    (Disease Target Structure Research Center, KRIBB)

  • Euijeon Woo

    (Disease Target Structure Research Center, KRIBB
    University of Science and Technology (UST))

  • Eun-Kyung Lim

    (Korea Research Institute of Bioscience and Biotechnology (KRIBB)
    KRIBB School of Biotechnology, UST
    Sungkyunkwan University (SKKU))

  • Juyeon Jung

    (Korea Research Institute of Bioscience and Biotechnology (KRIBB)
    Sungkyunkwan University (SKKU))

  • Taejoon Kang

    (Korea Research Institute of Bioscience and Biotechnology (KRIBB)
    Sungkyunkwan University (SKKU))

Abstract

Endonucleases have recently widely used in molecular diagnostics. Here, we report a strategy to exploit the properties of Argonaute (Ago) proteins for molecular diagnostics by introducing an artificial nucleic acid circuit with Ago protein (ANCA) method. The ANCA is designed to perform a continuous autocatalytic reaction through cross-catalytic cleavage of the Ago protein, enabling one-step, amplification-free, and isothermal DNA detection. Using the ANCA method, carbapenemase-producing Klebsiella pneumoniae (CPKP) are successfully detected without DNA extraction and amplification steps. In addition, we demonstrate the detection of carbapenem-resistant bacteria in human urine and blood samples using the method. We also demonstrate the direct identification of CPKP swabbed from surfaces using the ANCA method in conjunction with a three-dimensional nanopillar structure. Finally, the ANCA method is applied to detect CPKP in rectal swab specimens from infected patients, achieving sensitivity and specificity of 100% and 100%, respectively. The developed method can contribute to simple, rapid and accurate diagnosis of CPKP, which can help prevent nosocomial infections.

Suggested Citation

  • Hyowon Jang & Jayeon Song & Sunjoo Kim & Jung-Hyun Byun & Kyoung G. Lee & Kwang-Hyun Park & Euijeon Woo & Eun-Kyung Lim & Juyeon Jung & Taejoon Kang, 2023. "ANCA: artificial nucleic acid circuit with argonaute protein for one-step isothermal detection of antibiotic-resistant bacteria," Nature Communications, Nature, vol. 14(1), pages 1-13, December.
    • Handle: RePEc:nat:natcom:v:14:y:2023:i:1:d:10.1038_s41467-023-43899-4
    DOI: 10.1038/s41467-023-43899-4
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    References listed on IDEAS

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    1. Kevin Montagne & Guillaume Gines & Teruo Fujii & Yannick Rondelez, 2016. "Boosting functionality of synthetic DNA circuits with tailored deactivation," Nature Communications, Nature, vol. 7(1), pages 1-12, December.
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