Subtype screening of <i>bla</i><sub>IMP</sub> genes usin<i>g</i> bipartite primers for DNA sequencing<i> </i>

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  • Kawahara Ryuji
    Division of Microbiology, Osaka Institute of Public Health, Japan
  • Watahiki Masanori
    Department of Bacteriology, Toyama Institute of Health, Japan
  • Matsumoto Yuko
    Microbiological Testing and Research Division, Yokohama City Institute of Public Health, Japan
  • Uchida Kaoru
    Department of Bacteriology, Toyama Institute of Health, Japan
  • Noda Makiko
    Department of Infectious Diseases, Gifu Prefectural Research Institute for Health and Environmental Sciences, Japan
  • Masuda Kanako
    Hiroshima Prefectural Technology Research Institute, Public Health and Environment Center, Japan
  • Fukuda Chiemi
    Department of Microbiology, Kagawa Prefectural Research Institute for Environmental Sciences and Public Health, Japan
  • Abe Yuki
    Department of Microbiology, Ehime Prefectural Institute of Public Health and Environmental Science, Japan
  • Asano Yukiko
    Department of Microbiology, Ehime Prefectural Institute of Public Health and Environmental Science, Japan
  • Oishi Kazunori
    Department of Bacteriology, Toyama Institute of Health, Japan
  • Shibayama Keigo
    Department of Bacteriology II, National Institute of Infectious Diseases, Japan
  • Shinomiya Hiroto
    Department of Microbiology, Ehime Prefectural Institute of Public Health and Environmental Science, Japan

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Other Title
  • Subtype Screening of bla[IMP] Genes Using Bipartite Primers for DNA Sequencing
  • Subtype Screening of <i>bla</i><sub>IMP</sub> Genes Using Bipartite Primers for DNA Sequencing

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Abstract

<p>Genes conferring carbapenem resistance have spread worldwide among gram-negative bacteria. Subtyping of these genes has epidemiological value due to the global cross-border movement of people. Subtyping of blaIMP genes that frequently detected in Japan appears to be important in public health settings; however, there are few useful tools for this purpose. We developed a subtyping screening tool based on PCR direct sequencing, which targets the internal sequences of almost all blaIMP genes. The tool used bipartite multiplex primers with M13 universal sequences at the 5’-end. According to in silico analysis, among the 78 known IMP-type genes, except for blaIMP-81, 77 detected genes were estimated to be differentiated. In vitro evaluation indicated that sequences of amplicons of IMP-1, IMP-6, IMP-7, and IMP-20 templates were identical to their respective subtypes. Even if the amplicons were small or undetectable through the first PCR, sufficient amplicons for DNA sequencing were obtained through a second PCR using the M13 universal primers. In conclusion, our tool can be possibly used for subtype screening of blaIMP, which is useful for the surveillance of bacteria with blaIMP in clinical and public health settings or environmental fields.</p>

Journal

  • Japanese Journal of Infectious Diseases

    Japanese Journal of Infectious Diseases advpub (0), 592-599, 2021-03-31

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

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