Typing of Streptococcus mutans strains isolated from caries free and susceptible subjects by multilocus enzyme electrophoresis (original) (raw)
Related papers
Brazilian Archives of Biology and Technology, 2013
The objective of this study was to analyze the genotypic diversity, frequency of serotypes and the detection of mutacins from Streptococcus mutans isolates in caries-free and caries-active individuals. A total of 260 S. mutans isolated from 28 individuals with and without dental caries were subjected to AP-PCR and PCR screening of glucosyltransferase B, mutacin and serotype genes, which showed the presence of. 70 different genotypes. There was no statistically significant association between the presence of genes for serotypes and mutacins with dental caries. However, there was a statistically significant and a strong association between the higher genotypic diversity in the subjects with caries (r = 0.72, p = 0.001). There was an increase in the number of genotypes with increasing age (p <0.01).
Journal of Medical Microbiology, 2004
The present study evaluated the relationship between clonal diversity and some virulence traits of Streptococcus mutans isolated from eight caries-free and eight caries-active subjects. A total of 155 S. mutans isolates from caries-free subjects and 144 isolates from caries-active subjects were obtained from samples of saliva, dental plaque and tongue surface and identified by PCR. The isolates were submitted to arbitrarily primed (AP)-PCR (OPA-2 and OPA-13) and multilocus enzyme electrophoresis (MLEE) to establish the genotypic diversity. Production of water-insoluble glucan (WIG) (monitored by SDS-PAGE), final pH of cultures and the ability of bacterial cells to adhere to smooth glass in the presence of sucrose were measured. High and comparable abilities of MLEE and AP-PCR were found to distinguish S. mutans genotypes, using Simpson's index of discrimination (0 . 971 and 0 . 968, respectively). The results showed a significant difference (P , 0 . 01) in the number of genotypes when caries-free and caries-active groups were compared by both fingerprinting methods used. Final pH (P ¼ 0 . 32) and the percentage of adherence to a glass surface (P ¼ 0 . 62) did not show differences between the two groups; however, the intensities of WIG bands from the caries-active group were greater than those from the caries-free group (P , 0 . 01). In addition, WIG was positively correlated with the ability of S. mutans to adhere to a glass surface (r ¼ 0 . 34, P ¼ 0 . 02) from caries-active subjects. These data showed that AP-PCR analysis and MLEE are both effective methods for assessing the genetic relatedness of S. mutans. Using these techniques, it was found that there is a larger number of genotypes of S. mutans with increased ability to synthesize WIG in caries-active individuals.
Isolation and Typing of Streptococcus mutans and Streptococcus sobrinus from Caries-active Subjects
Contemporary clinical dentistry
Streptococcus mutans and Streptococcus sobrinus are main etiological agents of dental caries. The aim of the study was to isolate, identify, characterize, and determine the minimum inhibitory concentration (MIC) of S. mutans and S. sobrinus from caries-active subjects. Sixty-five plaque samples were collected from caries-active subjects aged between 35 and 44 years, processed and cultured on mitis salivarius bacitracin agar. All the bacterial isolates were subjected to morphotyping and the suspected colonies were identified by 16S rDNA sequencing. The S. mutans and S. sobrinus strains were characterized by biotyping and phylogenetic analysis. The MIC of ampicillin and erythromycin was determined by microtiter plate method. Of the study population, 41 isolates displayed typical colony morphologies of S. mutans and S. sobrinus. The 16S rDNA sequencing results revealed that 36 isolates were S. mutans and 5 isolates were S. sobrinus. The biotyping of these isolates demonstrated three bi...
Evaluation of genotypic diversity of Streptococcus mutans using distinct arbitrary primers
Journal of Applied Oral Science, 2008
treptococcus 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.
Characteristics of Clinical Isolates of Streptococcus mutans
Applied Sciences
Dental caries is an infectious disease which remains a significant health problem all over the world. The purpose of the study was to characterise a collection of 60 clinical isolates of S. mutans from adults’ and children’s dental plaque (natural biofilm). The paper describes the process of isolation, identification, analysis of biofilm formation and collection testing for the presence of 13 two-component systems (TCS) identified earlier in reference strain ATCC 700610 (UA159). In the case of S. mutans strains, plaque formation is specifically influenced by binary systems. All isolated strains of S. mutans form biofilm at high levels and possess a set of 26 genes forming TSC binary systems, which have an important role in plaque formation.
Brazilian Journal of Oral Sciences, 2014
Aim: To characterize the genetic variability of Streptococcus mutans isolates and to correlate this variability with different colonization profiles observed during dental caries in a sample of children. Methods: S. mutans samples were isolated from the saliva of 30 children with varying histories of dental caries, and they were characterized according to morphological and biochemical markers and the sequences of their 16S-23S intergenic spacer region. The genetic variability of the isolates was first assessed using Random Amplified Polymorphic DNA (RAPD) markers. Next, the isolates were differentiated by sequencing a specific region of the gene encoding the enzyme glucosyltransferase B (gtfB). Results: Characterization using RAPD markers uncovered significant genetic variability among the samples and indicated the existence of clusters, which allowed us to reconstruct both the origin and clinical history of the disease. By sequencing the 16S-23S intergenic region, it was found that all of the isolates belonged to the species S. mutans. Based on the genetic similarity of the isolates and pattern of amino acid variations identified by partial sequencing of the gtfB gene, base-pair changes were identified and correlated with different virulence patterns among the isolates. Conclusions: The partial sequencing of the gtfB gene can be a useful tool for elucidating the colonization patterns of S. mutans. As amino acid variations are likely to be correlated with differences in biological risk, molecular characterization, such as that described in this paper, could be the key for assessing the development of dental caries in children.
Iranian Journal of Basic Medical Sciences, 2013
Objective(s): The oral streptococci especially mutans Streptococci are related with the development of caries in humans. However, they are commonly distributed not only in populations with moderate or high caries incidence but also in populations having no or low caries experience. So, in this study, the differences between protein profiles of Streptococcus mutans from different sources were compared. Materials and Methods: Twelve S. mutans strains were isolated from caries-susceptible, caries-free subjects and also S. mutansATCC35668. Total proteins of the strains were extracted, and were analyzed through SDS-PAGE technique with coomassie staining. Results: The protein profiles of caries-susceptible subjects were different from those of caries-free subjects. The major protein bands of SDS-PAGE analysis were observed 26-100kDa and 45-57kDa in caries-susceptible and caries-free subjects, respectively. The major protein bands of SDS-PAGE analysis of S.mutans ATCC35668 were between 35-...
BMC oral health, 2017
Early childhood caries (ECC) is an aggressive condition that can affect teeth of young children. This study aimed to evaluate genotypic diversity and phenotypic traits of S. mutans isolated from dental biofilms of children with different caries status in comparison with caries free (CF) children. Streptococcus mutans strains were isolated from supragingival biofilm samples of CF, ECC and severe-ECC (S-ECC) children and genotyped by arbitrary-primer polymerase chain reaction - AP-PCR. S. mutans genotypes were tested for their ability to reduce the suspension pH through glycolysis, to tolerate extreme acid challenge and by their ability to form biofilm. Response variables were analyzed by ANOVA/Tukey or Kruskal-Wallis/Mann-Whitney tests at a 5% of significance. There was an increase in the prevalence of Streptococcus mutans in biofilms with the severity of dental caries. No differences in genotypic diversity and in acidogenicity of genotypes were found among CF, ECC and S-ECC children...