Single-Tube Multiplex Polymerase Chain Reaction for the Detection of Genes Encoding Enterobacteriaceae Carbapenemase

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  • Watahiki Masanori
    Department of Bacteriology, Toyama Institute of Health
  • Kawahara Ryuji
    Division of Microbiology, Osaka Institute of Public Health
  • Suzuki Masahiro
    Department of Microbiology and Medical Zoology, Aichi Prefectural Institute of Public Health Present Address: Department of Microbiology, School of Medicine, Fujita Health University
  • Aoki Miyako
    Department of Microbiology and Medical Zoology, Aichi Prefectural Institute of Public Health
  • Uchida Kaoru
    Department of Bacteriology, Toyama Institute of Health
  • Matsumoto Yuko
    Microbiological Testing and Research Division, Yokohama City Institute of Public Health
  • Kumagai Yuko
    Hygiene Division, Bacteriology Section, Akita Prefectural Research Center for Public Health and Environment
  • Noda Makiko
    Department of Infectious Diseases, Gifu Prefectural Research Institute for Health and Environmental Sciences
  • Masuda Kanako
    Hiroshima Prefectural Technology Research Institute, Public Health and Environment Center
  • Fukuda Chiemi
    Department of Microbiology, Kagawa Prefectural Research Institute for Environmental Sciences and Public Health
  • Harada Seiya
    Department of Microbiology, Kumamoto Prefectural Institute of Public Health and Environmental Science
  • Senba Keiko
    Department of Microbiology, Ehime Prefectural Institute of Public Health and Environmental Science
  • Suzuki Masato
    Antimicrobial Resistance Research Center, National Institute of Infectious Diseases
  • Matsui Mari
    Antimicrobial Resistance Research Center, National Institute of Infectious Diseases
  • Suzuki Satowa
    Antimicrobial Resistance Research Center, National Institute of Infectious Diseases
  • Shibayama Keigo
    Department of Bacteriology II, National Institute of Infectious Diseases
  • Shinomiya Hiroto
    Department of Microbiology, Ehime Prefectural Institute of Public Health and Environmental Science

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Other Title
  • Method : Single-Tube Multiplex Polymerase Chain Reaction for the Detection of Genes Encoding Enterobacteriaceae Carbapenemase

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Abstract

<p>A multiplex PCR assay in a single tube was developed for the detection of the carbapenemase genes of Enterobacteriaceae. Primers were designed to amplify the following six carbapenemase genes: blaKPC, blaIMP, blaNDM, blaVIM, blaOXA-48-like, and blaGES. Of 70 blaIMP variants, 67 subtypes were simulated to be PCR-positive based on in silico simulation and the primer-design strategy. After determining the optimal PCR conditions and performing in vitro assays, the performance of the PCR assay was evaluated using 51 and 91 clinical isolates with and without carbapenemase genes, respectively. In conclusion, the combination of multiplex PCR primers and QIAGEN Multiplex PCR Plus Kit was used to determine the best performance for the rapid and efficient screening of carbapenemase genes in Enterobacteriaceae. The assay had an overall sensitivity and specificity of 100%. This PCR assay compensates for the limitations of phenotypic testing, such as antimicrobial susceptibility testing and the modified carbapenem inactivation method, in clinical and public health settings.</p>

Journal

  • Japanese Journal of Infectious Diseases

    Japanese Journal of Infectious Diseases 73 (2), 166-172, 2020

    National Institute of Infectious Diseases, Japanese Journal of Infectious Diseases Editorial Committee

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