Real-time PCR for quantification of Streptococcus mutans

被引:69
|
作者
Yano, A
Kaneko, N
Ida, H
Yamaguchi, T
Hanada, N
机构
[1] Natl Inst Publ Hlth, Dept Oral Hlth, Shinjuku Ku, Tokyo 1628640, Japan
[2] Niigata Univ, Grad Sch Med & Dent Sci, Dept Oral Hlth Sci, Niigata 9518514, Japan
[3] BML Inc, Dept Bacteriol, Kawagoe, Saitama 3501101, Japan
[4] BML Inc, R&D Ctr, Div Clin Dev, Kawagoe, Saitama 3501101, Japan
关键词
real-time PCR; S; mutans; sobrinus; saliva; glucosyltransferase;
D O I
10.1111/j.1574-6968.2002.tb11451.x
中图分类号
Q93 [微生物学];
学科分类号
071005 ; 100705 ;
摘要
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. (C) 2002 Federation of European Microbiological Societies. Published by Elsevier Science B.V. All rights reserved.
引用
收藏
页码:23 / 30
页数:8
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