Development of Quantitative Real-Time PCR Primers for Detection of Streptococcus sobrinus.
10.11620/IJOB.2016.41.3.149
- Author:
Soon Nang PARK
1
;
Joong Ki KOOK
Author Information
1. Korean Collection for Oral Microbiology, Chosun University, 375 Seosuk-Dong, Dong-Gu, Gwangju 501-759, Republic of Korea. jkkook@chosun.ac.kr
- Publication Type:Original Article
- Keywords:
Streptococcus sobrinus;
rpoB;
qPCR primers
- MeSH:
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*
- From:International Journal of Oral Biology
2016;41(3):149-154
- CountryRepublic of Korea
- Language:Korean
-
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.
