Development of Quantitative Real-Time PCR Primers for Detection of Prevotella intermedia.
10.11620/IJOB.2015.40.4.205
- Author:
Soon Nang PARK
1
;
Joong Ki KOOK
Author Information
1. Korean Collection for Oral Microbiology, School of Dentistry, Chosun University, 375 Seosuk-Dong, Dong-Gu, Gwangju 501-759, Republic of Korea. jkkook@chosun.ac.kr
- Publication Type:Original Article
- Keywords:
Prevotella intermedia;
rpoB;
qPCR primers
- MeSH:
Bacteria;
Base Sequence;
Dental Plaque;
DNA;
DNA-Directed RNA Polymerases;
Polymerase Chain Reaction;
Prevotella intermedia*;
Prevotella*;
Real-Time Polymerase Chain Reaction*;
Sensitivity and Specificity
- From:International Journal of Oral Biology
2015;40(4):205-210
- CountryRepublic of Korea
- Language:Korean
-
Abstract:
Prevotella intermedia-specific quantitative real-time PCR (qPCR) primers were previously designed based on the nucleotide sequences of RNA polymerase beta-subunit gene (rpoB). However, the several clinical strains isolated from Korean populations are not detectable by the qPCR primers. The purpose of this study was to develop new P. intermedia-specific qPCR primers based on the rpoB. The specificity of the primers was determined by conventional PCR with 12 strains of P. intermedia and 52 strains (52 species) of non-P. intermedia bacteria. The sensitivity of primers was determined by qPCR with serial dilutions of the purified genomic DNAs (40 ng to 4 fg) of P. intermedia ATCC 25611T. The data indicated that only P. intermedia strains were detected by the P intermedia-specific qPCR primers (RTPiF2/RTPiR2); in addition, as little as 40 fg of P. intermedia genomic DNA could be detected. These results suggest that these qPCR primers are useful in detecting P. intermedia from the bacterial infectious lesions including dental plaque and oral tissue lesions.
