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Development of quantitative real-time PCR primers for detecting 42 oral bacterial species.

Korean Collection for Oral Microbiology and Department of Oral Biochemistry, School of Dentistry, Chosun University, 375 Seo-suk Dong, Dong-Gu, Gwangju, 501-759, Republic of Korea.
Archives of Microbiology (Impact Factor: 1.86). 05/2013; 195(7). DOI: 10.1007/s00203-013-0896-4
Source: PubMed

ABSTRACT In this study, we introduced species-specific quantitative real-time PCR (qPCR) primers designed based on a DNA-dependent RNA polymerase beta-subunit gene (rpoB) for detecting 42 oral bacterial species. The specificity of the qPCR primers was confirmed by conventional PCR with the genomic DNAs of 73-79 strains regarding 73-75 bacterial species including the type strain for the target species. The standard curves revealed the lower detection limits of 42 bacterial species-specific qPCR primers ranged from 4 to 40 fg below a cycle threshold (C T) value of 35, except Atopobium rimae, Fusobacterium nucleatum, Neisseria meningitidis, and Porphyromonas asaccharolytica which were 400 fg. These results suggest that 42 bacterial species-specific qPCR primers are suitable for applications in epidemiological studies related to oral infectious diseases such as periodontal diseases, endodontic infection, and dental caries.

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    ABSTRACT: In this study, we introduced species-specific quantitative real-time PCR (qPCR) primers designed based on a DNA-dependent RNA polymerase beta-subunit gene for detecting anginosus group streptococci (AGS), Streptococcus anginosus, S. constellatus, and S. intermedius. The specificity of the qPCR primers was confirmed by conventional PCR with the genomic DNAs of 76 strains regarding 44 bacterial species including the type strain for the target species. The standard curves revealed the lower detection limits of these species-specific qPCR primers was 40 fg at below a cycle threshold (CT) value of 35. These results suggest that AGS species-specific qPCR primers are suitable for applications in epidemiological studies associated with infectious diseases related to AGS.
    Archives of Microbiology 06/2014; 196(9). DOI:10.1007/s00203-014-1007-x · 1.86 Impact Factor

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May 20, 2014