Development of a Simple Method to Detect the Carbapenemase-Producing Genes blaNDM, blaOXA-48-like, blaIMP, blaKPC, and blaVIM Using a LAMP Method with Lateral Flow DNA Chromatography

Author:

Mikita Kei1ORCID,Tajima Moe1,Haque Anwarul2,Kato Yasuyuki2,Iwata Satoshi3,Suzuki Koichi4ORCID,Hasegawa Naoki1,Yano Hisakazu5,Matsumoto Tetsuya2

Affiliation:

1. Department of Infectious Diseases, Keio University School of Medicine, Tokyo 160-8582, Japan

2. Department of Infectious Diseases, Graduate School of Medicine, International University of Health and Welfare, Narita 286-8520, Japan

3. Department of Microbiology, Tokyo Medical University, Tokyo 160-8402, Japan

4. Department of Clinical Laboratory Science, Faculty of Medical Technology, Teikyo University, Tokyo 173-8606, Japan

5. Department of Microbiology and Infectious Diseases, Nara Medical University, Nara 634-8522, Japan

Abstract

Infections by carbapenemase-producing Enterobacterales constitute a global public health threat. The rapid and efficient diagnosis of Enterobacterales infection is critical for prompt treatment and infection control, especially in hospital settings. We developed a novel loop-mediated isothermal amplification (LAMP) method combined with DNA chromatography to identify five major groups of carbapenemase-producing genes (blaNDM, blaOXA-48-like, blaIMP, blaKPC, and blaVIM). This method uses DNA–DNA hybridization-based detection in which LAMP products can be easily visualized as colored lines. No specific technical expertise, expensive equipment, or special facilities are required for this method, allowing its broad application. Here, 73 bacteria collections including strains with carbapenemase-producing genes were tested. Compared to sequencing results, LAMP DNA chromatography for five carbapenemase-producing genes had a sensitivity and specificity of 100% and >97%, respectively. This newly developed method can be a valuable rapid diagnostic test to guide appropriate treatments and infection control measures, especially in resource-limited settings.

Funder

Japan Agency for Medical Research and Development

JSPS KAKENHI

Publisher

MDPI AG

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