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Journal of Clinical Microbiology, June 2005, p. 2948-2951, Vol. 43, No. 6
0095-1137/05/$08.00+0     doi:10.1128/JCM.43.6.2948-2951.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.

Use of the Genomic Subtractive Hybridization Technique To Develop a Real-Time PCR Assay for Quantitative Detection of Prevotella spp. in Oral Biofilm Samples

Division of Community Oral Health Science, Kyushu Dental College, Kitakyushu 803-8580,¹ Department of Oral Bacteriology, Ohu University School of Dentistry, Koriyama 963-8611, Japan²

Received 17 December 2004/ Returned for modification 31 January 2005/ Accepted 10 February 2005

Genomic subtractive hybridization was used to design Prevotella nigrescens-specific primers and TaqMan probes. Based on this technique, a TaqMan real-time PCR assay was developed for quantifying four oral black-pigmented Prevotella species. The combination of real-time PCR and genomic subtractive hybridization is useful for preparing species-specific primer-probe sets for closely related species.