Int J Oral Biol.  2016 Sep;41(3):149-154. 10.11620/IJOB.2016.41.3.149.

Development of Quantitative Real-Time PCR Primers for Detection of Streptococcus sobrinus

• Park SN
• Kook JK

• KMID: 2354203
• DOI: http://doi.org/10.11620/IJOB.2016.41.3.149

Abstract

The purpose of this study was to develop Streptococcus sobrinus-specific qPCR primers based on the nucleotide sequence of the RNA polymerase β-subunit gene (rpoB). The specificity of the primers was determined by conventional polymerase chain reaction (PCR) with 12 strains of S. sobrinus and 50 strains (50 species) of non-S. sobrinus bacteria. The sensitivity of the primers was determined by quantitative real-time PCR (qPCR) with serial dilutions of the purified genomic DNAs (40 ng to 4 fg) of S. sobrinus ATCC 33478(T). The specificity data showed that the S. sobrinus-specific qPCR primers (RTSsob-F4/RTSsob-R4) detected only the genomic DNAs of S. sobrinus strains with a detection limit of up to 4 fg of S. sobrinus genomic DNA. Our results suggest that the RTSsob-F4/RTSsob-R4 primers are useful in detecting S. sobrinus with high sensitivity and specificity for epidemiological studies of dental caries.

Keyword

Streptococcus sobrinus; rpoB; qPCR primers

MeSH Terms

Bacteria
Base Sequence
Dental Caries
DNA
DNA-Directed RNA Polymerases
Epidemiologic Studies
Limit of Detection
Polymerase Chain Reaction
Real-Time Polymerase Chain Reaction*
Sensitivity and Specificity
Streptococcus sobrinus*
Streptococcus*
DNA
DNA-Directed RNA Polymerases