Analysis of total oral microorganisms in saliva using real-time PCR and colony forming unit

Su-Min Yoo   Seong-Kug Jeong1   Hyun-Jun Yoo2   Jong-Hwa Jang3,*   

Departments of Dental Hygiene, Kyungdong University
1Departments of Preventive Dentistry, School of Dentistry, Chosun University
2Departments of Preventive Dentistry, School of Dentistry, Dankuk University
3Departments of Dental Hygiene, Hanseo University

Abstract

Objectives: The purpose of this study was to compare colony forming unit (CFU) method and multiplex real-time polymerase chain reaction (MRT-PCR) method for accurate quantitative analysis of bacteria. Methods: We compared the CFU method and the MRT-PCR method, which are still used in Korea, for Prevotella intermedius (P. intermedius), a periodontal disease pathogen selected by MRT-PCR, and Streptococcus mutans (S. mutans), a dental caries causative organism. The subjects of this study were 30 patients who visited the C dental hospital. Results: Total microorganisms in MRT-PCR method were significantly higher in both types of bacteria (p<0.05), since DNA of dead bacteria was also analyzed. This was because the periodontal dise(-) anaerobes, and even dead bacteria contain large amounts of toxic substances called LPS in the extracellular membrane, and fimbriae and pili, which are motility structures, still remain as a strong toxic substance in periodontal tissue. Conclusions: Therefore, in terms of the total amount of bacteria found, the MRT-PCR method will be a useful technique for searching all the bacteria in the oral cavity including live bacteria, as well as sterilization.

Acknowledgements

이 논문은 2014년도 정부(미래창조과학부)의 재원으로 한국연구재단의 지원을 받아 수행된 기초연구사업임(2014R1A1A3051084).

Figures & Tables

Table 1. Primer and probe used for multiplex real-time PCR