Sato T, Matsuyama J, Kumagai T, Mayanagi G, Yamaura M, Washio J, Takahashi N
Division of Oral Ecology and Biochemistry, Tohoku University Graduate School of Dentistry, Sendai, Japan.
Lett Appl Microbiol. 2003;37(1):66-9. doi: 10.1046/j.1472-765x.2003.01359.x.
Mutans streptococci such as Streptococcus mutans and Streptococcus sobrinus have been implicated in human dental caries. In an attempt to develop a rapid and sensitive method for detecting Strep. mutans and Strep. sobrinus in dental plaque, a nested PCR amplification based on the 16S rRNA gene was employed.
A universal set of PCR primers for bacterial 16S rRNA gene was introduced for the first PCR, and then two sets of primers specific for the 16S rRNA gene sequences of either Strep. mutans or Strep. sobrinus were used for the second PCR. Eighteen plaque samples were analyzed, and a nested PCR was shown to be more sensitive for detecting Strep. mutans and Strep. sobrinus than direct PCR.
CONCLUSIONS, SIGNIFICANCE AND IMPACT OF THE STUDY: The 16S rRNA gene-based nested PCR method is a rapid and sensitive method for the detection of mutans streptococci, and may also be suitable for carrying out large-scale studies on the cariogenicity of mutans streptococci.
变形链球菌和远缘链球菌等变形链球菌与人类龋齿有关。为了开发一种快速、灵敏的方法来检测牙菌斑中的变形链球菌和远缘链球菌,采用了基于16S rRNA基因的巢式PCR扩增技术。
第一次PCR采用一套通用的细菌16S rRNA基因PCR引物,然后第二次PCR使用两套分别针对变形链球菌或远缘链球菌16S rRNA基因序列的引物。对18个菌斑样本进行了分析,结果表明巢式PCR检测变形链球菌和远缘链球菌比直接PCR更灵敏。
研究的结论、意义和影响:基于16S rRNA基因的巢式PCR方法是一种检测变形链球菌的快速、灵敏方法,也可能适用于对变形链球菌致龋性进行大规模研究。