Tracing genotypes of mutans streptococci on tooth sites by random amplified polymorphic DNA (RAPD) analysis

The aim of this study was to clarify the distribution and persistence of mutans streptococci on different tooth sites in the same oral cavity. Thirteen subjects, aged 20-40 years, were examined. Salivary levels of mutans streptococci, caries prevalence, oral hygiene habits and status of tooth surfaces sampled were recorded. Plaque samples were obtained from four sites, the mesial and buccal surfaces of the first permanent molar on the right side of the lower jaw (46m and 46b), the distal surface of the first permanent premolar (24d) and the mesial surface of the second permanent premolar (25m) on the left side of the upper jaw, using sterile toothpicks on two occasions at 4-7-month intervals. The samples were cultivated on site-specific Strip mutans. Up to 10 colonies/site were isolated when present and genotyped by random amplified polymorphic DNA (RAPD) analysis, after species identification with PCR. Genotyping was also performed by restriction endonuclease analysis (REA) on 148 isolates, and results were consistent with the RAPD results. Most mutans streptococcus-positive samples were obtained from 46m. Within each individual, the same genotype occurred on at least two sites on all but one sampling occasion. A maximum of seven different genotypes were found in an individual. For a particular tooth site, four genotypes occurred simultaneously and taking both sampling occasions together the maximum was six different types. The same genotypes/types were found again after 4-7 months on 25 sites in 12 subjects. Fifteen sites were mutans streptococcus-positive on only one sampling occasion. The results indicate that several different genotypes of mutans streptococci colonize a tooth site, and that the same genotype colonizes several sites in the same oral cavity. Persistence of genotypes on a site for several months and interindividual differences in the occurrence of genotypes were also found

Evaluation of genotypic diversity of Streptococcus mutans using distinct arbitrary primers

Streptococcus mutans has been considered one of the main etiological agents of dental caries and the genotypic diversity rather than its salivary counts may be considered as a virulence factor of this bacterium. For genotyping with polymerase chain reaction (PCR) with arbitrary primers, several primers have been used in order to improve complexity and specificity of amplicon patterns. Thus, the aim of this study was to evaluate the degree of agreement of genotypic identification among AP-PCR reactions performed with 5 distinct arbitrary primers of S. mutans isolated from saliva. Stimulated saliva was collected from 11 adult volunteers for isolation of S. mutans, and a total of 88 isolates were genotyped with arbitrary primers OPA 02, 03, 05, 13 and 18. Fourteen distinct genotypes were identified in the saliva samples. Most volunteers (9 out of 11) presented only one genotype. The results of the present study suggest that primers OPA 02, 03, 05 and 13 were suitable for genotypic identification of S. mutans isolates of saliva from adult volunteers

Risk factors for early colonization of mutans streptococci - a multiple logistic regression analysis in Swedish 1-year-olds

Background: Mutans streptococci (MS) are closely related to the development of dental caries and are usually established in the oral cavity during early childhood. The aim of the study was to identify factors associated with the presence of MS in Swedish 1-year-olds. Methods: Parents completed a questionnaire on different caries-associated factors and an oral bacterial sample was collected from 1,050 (526 boys, 524 girls) 1-year-olds. Multiple logistic regression analyses were performed to identify risk factors for colonization with MS. Results: MS were found in 27% of the 1-year-olds with teeth. High or very high MS scores (2-3) were found in 72 (7%) of the children. MS score was correlated to the number of erupted teeth (p less than 0.001). No difference due to gender was found. Multiple logistic regression analysis showed that presence of bacteria was associated with: caries in a sibling, other beverages than water between meals, and more than 8 erupted teeth. High or very high MS scores (2-3) were associated with other beverages than water between meals, and more than 8 erupted teeth. Conclusions: Number of teeth present, diet and family aspects were factors associated with presence of MS in 1-year-olds. To develop high or very high MS scores, the number of erupted teeth and dietary habits are important.Funding Agencies|Public Dental Service/Folktandvarden Ostergotland, Sweden</p