Passive Filtration, Rapid Scanning Electron Microscopy, and Matrix-Assisted Laser Desorption Ionization-Time of Flight Mass Spectrometry for Treponema Culture and Identification from the Oral Cavity

J Clin Microbiol. 2019 Sep 24;57(10):e00517-19. doi: 10.1128/JCM.00517-19. Print 2019 Oct.

Abstract

We present here a new passive-filtration-based culture device combined with rapid identification with a new electron microscope (Hitachi TM4000) for the detection and culture of Treponema species from the human oral cavity. Of the 44 oral samples cultivated, 15 (34%) were found to be positive for Treponema using electron microscopy and were also culture positive. All were subcultured on agar plates; based on genome sequencing and analyses, 10 were strains of Treponema pectinovorum and 5 were strains of Treponema denticola The 29 samples that were negative for Treponema remained culture negative. In addition, 14 Treponema species ordered from the DSMZ collection were cultured in the T-Raoult culture medium optimized here. Finally, matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) was used and 30 novel spectra were added to the MALDI-TOF MS database. We have successfully developed a new and effective method for treponemal detection, culture, and identification.

Keywords: Treponema; culture; microscopy; oral cavity; passive filtration.

Publication types

  • Research Support, Non-U.S. Gov't
  • Retracted Publication

MeSH terms

  • Bacterial Typing Techniques / methods
  • Bacterial Typing Techniques / standards
  • Filtration*
  • Genome, Bacterial
  • Humans
  • Microscopy, Electron, Scanning*
  • Mouth / microbiology*
  • Phylogeny
  • RNA, Ribosomal, 16S
  • Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization*
  • Treponema / classification*
  • Treponema / genetics
  • Treponemal Infections / diagnosis*
  • Treponemal Infections / microbiology*

Substances

  • RNA, Ribosomal, 16S