PCR detection of periodontal bacteria and Streptococcus mutans in the biological samples from the oral cavity

• Published in: Klinicheskaia laboratornaia diagnostika no. 4; p. 35
• Main Authors: Chukhlovin, A B, Solov'eva, A M, Matelo, S K, Dmitriev, A V, Morozova, E B, Bobrov, A P, Totolian, A A
• Format: Journal Article
• Language: Russian
• Published: Russia (Federation) 01-04-2007
• Subjects: Adolescent; Adult; Aged; Aged, 80 and over; Aggregatibacter actinomycetemcomitans - genetics; Aggregatibacter actinomycetemcomitans - isolation & purification; Bacteroides - genetics; Bacteroides - isolation & purification; Child; Child, Preschool; Colony Count, Microbial; DNA, Bacterial - analysis; DNA, Bacterial - genetics; Female; Humans; Male; Middle Aged; Mouth - microbiology; Polymerase Chain Reaction - methods; Porphyromonas gingivalis - genetics; Porphyromonas gingivalis - isolation & purification; Sensitivity and Specificity; Stomatognathic Diseases - genetics; Stomatognathic Diseases - microbiology; Streptococcus mutans - genetics; Streptococcus mutans - isolation & purification
• ISSN: 0869-2084

The paper describes an optimized technique for DNA isolation from the Streptococcus mutans cultures and from the clinical samples including proteinase K treatment and detergent lysis, followed by sorbent-based enrichment. This technique was employed for isolating DNA from the periodontal bacteria Porphyromonas gingivalis, Actinobacillus actinomycetemcomitans, and Bacteroides forsythus. A multiplex PCR technique was adapted for large-scale studies of clinical samples. An original PCR method was developed for the semiquantitative detection of Str. mutans, showing the sensitivity of 100 genome copies per reaction. Extensive studies of approximately 2,000 individuals have demonstrated that these methods are applicable to the study of the dental and gingival microflora in the biological sample from the oral cavity of persons with dental diseases from various age groups.