Restriction fragment-length polymorphism analysis of 16S ribosomal RNA genes amplified by polymerase chain reaction for rapid identification of cultivable oral treponemes

T. Sato, Howard K. Kuramitsu

Research output: Contribution to journalArticle

9 Citations (Scopus)

Abstract

Although oral treponemes are among the most frequently found bacteria in periodontal Pockets, identification of these organisms can be difficult. In this study, restriction fragment-length polymorphism (RFLP) analysis of polymerase chain reaction (PCR)-amplified 16S ribosomal RNA genes (16S rRNA gene PCR-RFLP) was used to generate restriction profiles of reference strains of oral treponemes including Treponema denticola, Treponema socranskii, Treponema vincentii, Treponema pectinovorum and Treponema medium as well as for Treponema phagedenis and Treponema pallidum and five treponeme strains isolated from human periodontal pockets. Before RFLP analysis, the 16S rRNA gene sequences were obtained from the GenBank database, and the analysis of the theoretical banding patterns for HpaII suggested good species discrimination. 16S rRNA gene sequences were amplified from isolated genomic DNA samples by PCR with spirochete-specific primers. The PCR products were then purified and characterized by single digestion with restriction endonuclease HpaII, and this allowed discrimination between the respective reference strains. Five clinical isolates, four T. denticola and one T. socranskii, were assigned on the basis of their restriction profiles by digestion with HpaII. 16S rRNA gene PCR-RFLP using HpaII is a rapid and reliable method for differentiation of cultivable oral treponemes.

Original languageEnglish
Pages (from-to)117-121
Number of pages5
JournalOral Microbiology and Immunology
Volume14
Issue number2
DOIs
Publication statusPublished - 1999 Apr

Keywords

  • 16S rRNA
  • Oral treponeme
  • PCR-RFLP

ASJC Scopus subject areas

  • Microbiology
  • Immunology
  • Dentistry(all)
  • Microbiology (medical)

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