Differentiation of Streptococcus mutans and Streptococcus sobrinus via genotypic and phenotypic profiles from three different populations.

Routine identification of Streptococcus mutans and Streptococcus sobrinus is generally based upon growth on various selective media, colony morphology and biochemical characteristics. We examined various approaches of differentiating these two species through a combination of the conventional phenotypic methodology with chromosomal DNA fingerprint (CDF) and arbitrarily primed polymerase chain reaction (AP-PCR) methods. Initially, ten ATCC type strains and 20 randomly selected clinical isolates of mutans streptococci (MS) were characterized and grouped into two major types based on patterns generated by the CDF using HaeIII digestion. The CDF's patterns with restriction fragments equal to or greater than 6.6 kb were defined as the CDF-1 group. The CDF's patterns with restriction fragments less than 6.6 kb were defined as the CDF-2 group. Both groups were then examined for biotype, serotype, and composition of DNA via thermal denaturation. AP-PCR was applied and evaluated for the capability of delineating S. mutans from S. sobrinus strains. Results of this study showed that all CDF-1 strains fit within a G+C range of 36.2% to 42.2%, whereas the CDF-2 strains had a G+C range of 45.8% to 47.0%. The serotyping assay exhibited 100% sensitivity, 90% specificity and 86.7% agreement with the CDF. The biotyping assay presented the poorest specificity (38.5%), indicating the highest variability. The capability of AP-PCR in differentiation of S. mutans from S. sobrinus was comparable to the CDF method, suggesting that either of these two approaches can and may serve as a viable alternative method to serotyping or biotyping of MS.

Genotypes of mutans streptococci tend to persist in their host for several years.

The aim of this study was to evaluate the consistency of the prevalence of mutans streptococci in a group of Swedish families. Eleven families, which had previously been examined for genotypes of mutans streptococci, were re-examined after 2-5 years. The families consisted of mother, father and a child (mean age 7.2 years at the follow-up examination). One father did not participate. Pooled plaque samples were obtained from buccal and occlusal surfaces. Isolates of mutans streptococci were genotyped using chromosomal DNA digested with restriction endonuclease HaeIII, separated by gel electrophoresis and visualised through UV illumination after ethidium bromide staining. Comparing the DNA fingerprints of mutans streptococci found at baseline and follow-up, 9 children harboured one or two genotypes which were similar on the two sampling occasions. Two of these children had also gained a genotype. The remaining 2 children had lost a genotype each and 1 of them had gained two new genotypes. All 21 adults showed one or two genotypes identical to those found at baseline. Nine of these 21 adults had also lost one genotype. Four of these 9 and additionally 4 of the remaining adults showed one or two new genotypes. Six mother-child pairs shared a genotype at baseline and this pattern remained for five pairs at the end of the study. The results suggest that genotypes of mutans streptococci have a fairly high degree of consistency in children between 3 and 8 years of age as well as in adults, indicating persistence of the strains. However, the results also indicate that some subjects may gain and/or lose genotypes.

Genotyping shows different strains of mutans streptococci between father and child and within parental pairs in Swedish families.

The purpose of this study was to investigate whether there was an intrafamilial similarity of mutans streptococcal strains in some Swedish families using chromosomal DNA fingerprinting. Plaque samples were obtained from buccal and occlusal surfaces of 25 three-year-old children, their mothers and 18 fathers. The colonization levels of mutants streptococci were estimated with the "Strip mutans" test, and caries experience was scored by decayed, missing and filled teeth or decayed, extracted and filled teeth. Interviews about medical history, diet regimes, breastfeeding and care of the child were performed. In 11 families isolates of mutans streptococci were detected in all three individuals. These isolates were serotyped by immunofluorescent technique and genotyped using the restriction endonuclease Hae III. The results showed that 5 children harbored mutans streptococci genotypes different from their parents. Six children showed genotypes identical to their mothers. None of the children harbored genotypes similar to their fathers, even though two thirds of the fathers had high or very high mutans streptococci levels. No matching of genotypes was observed within the 11 parental pairs. Mothers as primary caregivers with high "Strip mutans" scores were more often observed in the group with identical genotypes within the mother-child pairs, the "matching group", than in the "no-matching group". These data indicate that the fathers and the children had not acquired each others' strains of mutans streptococci nor had the spouses. The results suggest that the children acquired mutans streptococci both from outside and inside the family.