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

Hyowon Jang, Jayeon Song, Sunjoo Kim, Jung-Hyun Byun, Kyoung G. Lee, Kwang-Hyun Park, Euijeon Woo, Eun-Kyung Lim, Juyeon Jung and Taejoon Kang ()
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Hyowon Jang: Korea Research Institute of Bioscience and Biotechnology (KRIBB)
Jayeon Song: Korea Research Institute of Bioscience and Biotechnology (KRIBB)
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
Eun-Kyung Lim: Korea Research Institute of Bioscience and Biotechnology (KRIBB)
Juyeon Jung: Korea Research Institute of Bioscience and Biotechnology (KRIBB)
Taejoon Kang: Korea Research Institute of Bioscience and Biotechnology (KRIBB)

Nature Communications, 2023, vol. 14, issue 1, 1-13

Abstract: 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.

Date: 2023
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DOI: 10.1038/s41467-023-43899-4

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