Rapid Identification of Mycoplasma pulmonis Isolated from Laboratory Mice and Rats Using Matrix-Assisted Laser Desorption Ionization Time-of-Flight Mass Spectrometry

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  • GOTO Kazuo
    Department of Clinical Laboratory Medicine, School of Medical Technology, Teikyo University Graduate School of Medical Technology, Teikyo University Department of Clinical Laboratory Medicine, School of Medical Technology, Teikyo University, 2–11–1 Kaga, Itabashi-ku, Tokyo 173–8605, Japan Graduate School of Medical Technology, Teikyo University, 2–11–1 Kaga, Itabashi-ku, Tokyo 173–8605, Japan
  • YAMAMOTO Mikachi
    Laboratory of Space and Environmental Medicine, Graduate School of Medical Technology, Teikyo University Laboratory of Space and Environmental Medicine, Graduate School of Medical Technology, Teikyo University, 2–11–1 Kaga, Itabashi-ku, Tokyo 173–8605, Japan
  • ASAHARA Miwa
    Graduate School of Medical Technology, Teikyo University Graduate School of Medical Technology, Teikyo University, 2–11–1 Kaga, Itabashi-ku, Tokyo 173–8605, Japan
  • TAMURA Takashi
    Graduate School of Medical Technology, Teikyo University Graduate School of Medical Technology, Teikyo University, 2–11–1 Kaga, Itabashi-ku, Tokyo 173–8605, Japan
  • MATSUMURA Mitsuru
    Department of Clinical Laboratory Medicine, School of Medical Technology, Teikyo University Graduate School of Medical Technology, Teikyo University Department of Clinical Laboratory Medicine, School of Medical Technology, Teikyo University, 2–11–1 Kaga, Itabashi-ku, Tokyo 173–8605, Japan Graduate School of Medical Technology, Teikyo University, 2–11–1 Kaga, Itabashi-ku, Tokyo 173–8605, Japan
  • HAYASHIMOTO Nobuhito
    Central Institute for Experimental Animals, ICLAS Monitoring Center Central Institute for Experimental Animals, ICLAS Monitoring Center, 1430 Nogawa, Miyamae-ku, Kawasaki, Kanagawa 216–0001, Japan
  • MAKIMURA Koichi
    Graduate School of Medical Technology, Teikyo University Laboratory of Space and Environmental Medicine, Graduate School of Medical Technology, Teikyo University Graduate School of Medical Technology, Teikyo University, 2–11–1 Kaga, Itabashi-ku, Tokyo 173–8605, Japan Laboratory of Space and Environmental Medicine, Graduate School of Medical Technology, Teikyo University, 2–11–1 Kaga, Itabashi-ku, Tokyo 173–8605, Japan

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  • Rapid Identification of <i>Mycoplasma pulmonis</i> Isolated from Laboratory Mice and Rats Using Matrix-Assisted Laser Desorption Ionization Time-of-Flight Mass Spectrometry

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Mycoplasma species identification is based on biochemical, immunological, and molecular methods that require several days for accurate identification. Matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS) is a novel method for identification of bacteria and has recently been introduced into the clinical microbiology laboratory as a rapid and accurate technique. This method allows a characteristic mass spectral fingerprint to be obtained from whole inactivated mycoplasmal cells. In this study, we evaluated the performance of the MALDI-TOF MS for the identification of Mycoplasma by comparison with standard sequence analysis of 16S rRNA. We developed the first database of MALDI-TOF MS profiles of Mycoplasma species, containing Mycoplasma pulmonis, M. arthritidis, and M. neurolyticum, which are the most common pathogens in mice and/or rats, and species-specific spectra were recorded. Using the database, 6 clinical isolates were identified. Six tracheal swabs from 4 mice and 2 rats were cultured on PPLO agar for 4 to 7 days, and the colonies were directly applied to analyze the protein profiles. Five strains were identified as M. pulmonis, and 1 strain from a mouse was identified as M. neurolyticum (spectral scores were >2.00); the results were consistent with the results of the 16S rRNA gene sequence analysis (homologies>97.0%). These data indicate that MALDI-TOF MS can be used as a clearly rapid, accurate, and cost-effective method for the identification of M. pulmonis isolates, and this system may represent a serious alternative for clinical laboratories to identify Mycoplasma species.

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