Simple and rapid detection of Streptococcus mutans and Streptococcus sobrinus in human saliva by polymerase chain reaction

T. Oho, Yoshihisa Yamashita, Y. Shimazaki, M. Kushiyama, Toshihiko Koga

Research output: Contribution to journalArticle

124 Citations (Scopus)

Abstract

Streptococcus mutans and Streptococcus sobrinus are major pathogens causing dental caries in humans. A simple and rapid method to detect these species in human saliva simultaneously was developed using the polymerase chain reaction (PCR). Chromosomal DNA was extracted by boiling bacterial cells in lysis solution containing 1% Triton X-100. Oligonucleotide primers specific for portions of the glucosyltransferase genes (gtfB of S. mutans and gtfI of S. sobrinus) were designed. After PCR using two sets of these primers, S. mutans and S. sobrinus were specifically identified. The method was capable of amplifying DNA fragments specific for these species from chromosomal DNA extracted from 1 X 103 cells, or from μl of clinical saliva samples containing 1 X 103 colony-forming units of either streptococcal species. A second PCR, using the first PCR product as a template with newly designed internal primers, made it possible to detect 1 x 102 colony-forming units of either streptococcal species in 10 μl of saliva samples. These results indicate that the PCR method developed in this study is useful for detecting S. mutans and S. sobrinus in saliva and that it can be used in epidemiological studies to evaluate the prevalence level of these organisms.

Original languageEnglish
Pages (from-to)258-262
Number of pages5
JournalOral Microbiology and Immunology
Volume15
Issue number4
DOIs
Publication statusPublished - Aug 2000

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All Science Journal Classification (ASJC) codes

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

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