Simple and rapid detection of <i>Streptococcus mutans</i> and <i>Streptococcus sobrinus</i> in human saliva by polymerase chain reaction

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<jats:p> <jats:italic>Streptococcus mutans</jats:italic> and <jats:italic>Streptococcus sobrinus</jats:italic> 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 (<jats:italic>gtfB</jats:italic> of <jats:italic>S. mutans</jats:italic> and <jats:italic>gtfI</jats:italic> of <jats:italic>S. sobrinus</jats:italic>) were designed. After PCR using two sets of these primers, <jats:italic>S. mutans</jats:italic> and <jats:italic>S. sobrinus</jats:italic> were specifically identified. The method was capable of amplifying DNA fragments specific for these species from chromosomal DNA extracted from 1×10<jats:sup>3</jats:sup> cells, or from 10 μl of clinical saliva samples containing 1×10<jats:sup>3</jats:sup> 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 1×10<jats:sup>2</jats:sup> 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 <jats:italic>S. mutans</jats:italic> and <jats:italic>S. sobrinus</jats:italic> in saliva and that it can be used in epidemiological studies to evaluate the prevalence level of these organisms.</jats:p>

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