Yano Akira, Kaneko Noboru, Ida Hirohisa, Yamaguchi Toshikazu, Hanada Nobuhiro
Department of Oral Health, National Institute of Public Health, 1-23-1 Toyama, Shinjuku-ku, 162-8640, Tokyo, Japan.
FEMS Microbiol Lett. 2002 Nov 19;217(1):23-30. doi: 10.1111/j.1574-6968.2002.tb11451.x.
A real-time polymerase chain reaction (PCR) assay was developed for the quantification of Streptococcus mutans. Primers targeting gtf genes of S. mutans were designed and tested for their specificity using 28 oral streptococcal strains, three other bacterial strains, and human DNA. The primers could amplify specifically the target DNA fragment from a mixture of oral streptococcus genomic DNA containing about 10 fg to 10 ng of S. mutans genome DNA. The real-time PCR produced a linear quantitative detection range over concentrations spanning seven exponential values, with a detection limit of a few copies of S. mutans' genomic DNA per reaction tube. The results of the real-time PCR assay corresponded well to those of conventional culture assays for S. mutans in saliva samples. A real-time PCR assay for Streptococcus sobrinus and Streptococcus downei was also established and produced results that corresponded well to those from conventional culture assays for S. sobrinus in saliva samples. These assays will be useful as a new means to assess one of the important risk factors for caries.
开发了一种用于定量变形链球菌的实时聚合酶链反应（PCR）检测方法。设计了针对变形链球菌gtf基因的引物，并使用28种口腔链球菌菌株、其他三种细菌菌株和人类DNA对其特异性进行了测试。这些引物能够从含有约10 fg至10 ng变形链球菌基因组DNA的口腔链球菌基因组DNA混合物中特异性扩增目标DNA片段。实时PCR在跨越七个指数值的浓度范围内产生线性定量检测范围，每个反应管中变形链球菌基因组DNA的检测限为几份拷贝。实时PCR检测结果与唾液样本中变形链球菌的传统培养检测结果非常吻合。还建立了针对远缘链球菌和唐氏链球菌的实时PCR检测方法，其结果与唾液样本中远缘链球菌的传统培养检测结果非常吻合。这些检测方法将作为评估龋齿重要风险因素之一的新手段。
