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Rapid determination of carbapenem resistance by low-cost colorimetric methods: Propidium Iodide and alamar blue staining

  • Microbial Ecology and Environmental Microbiology
  • Published:
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Abstract

Carbapenems are a class of β-lactam antibiotics with a broad antimicrobial activity spectrum. Owing to their sturdy structures resistant to most β-lactamases, they have been regarded as one of the last-resort antibiotics for combating multidrugresistant bacterial infections. However, the emergence of carbapenem resistance increases predominantly in nosocomial pathogens. To prevent spread of carbapenem resistance in early stages, it is imperative to develop rapid diagnostic tests that will substantially reduce the time and cost in determining carbapenem resistance. Thus, we devised a staining-based diagnostic method applicable to three different Gram-negative pathogens of Acinetobacter baumannii, Escherichia coli, and Klebsiella pneumoniae, all with the high potential to develop carbapenem resistance. Regardless of the resistance mechanisms presented by bacterial species and strains, double staining with propidium iodide (PI) and alamar blue (AB) identified resistant bacteria with an average sensitivity of 95.35%, 7 h after imipenem treatments in 343 clinical isolates. Among the three species tested, A. baumannii showed the highest diagnostic sensitivity of 98.46%. The PI and ABmediated staining method could be a promising diagnostic method with high-throughput efficacy and low cost.

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Acknowledgments

This research was supported by a grant (HI15C1733) from the Korea Health Industry Development Institute (KHIDI), funded by the Ministry of Health & Welfare; a grant from Priority Research Centers Program (2019R1A6A1A11051471), funded by the National Research Foundation of Korea (NRF); and a grant (20162MFDS010) from Ministry of Food and Drug Safety in 2020.

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    Authors and Affiliations

    1. Department of Molecular Science and Technology, Ajou University, Suwon, 16499, Republic of Korea

      Jiyoon Choi, Jiwon Baek & Hyunjin Yoon

    2. Department of Genetic Engineering and Center for Human Interface Nano Technology, Sungkyunkwan University, Suwon, 16419, Republic of Korea

      Daehyuk Kweon

    3. Department of Molecular Cell Biology, Sungkyunkwan University, School of Medicine, Suwon, 16419, Republic of Korea

      Kwan Soo Ko

    4. Asia-Pacific Research Foundation for Infectious Diseases, Seoul, 06367, Republic of Korea

      Kwan Soo Ko

    5. Department of Applied Chemistry and Biological Engineering, Ajou University, Suwon, 16499, Republic of Korea

      Hyunjin Yoon

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    1. Jiyoon Choi
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    Correspondence to Hyunjin Yoon.

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    Choi, J., Baek, J., Kweon, D. et al. Rapid determination of carbapenem resistance by low-cost colorimetric methods: Propidium Iodide and alamar blue staining. J Microbiol. 58, 415–421 (2020). https://doi.org/10.1007/s12275-020-9549-x

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