Identification of oral mitis group streptococci by arbitrarily primed polymerase chain reaction

Abstract

"Mitis group" streptococci are commensal but may play some role in dental caries, septicemia or endocarditis. Rapid genotypic identification would aid studies of dental plaque ecology, or diagnostic use. AP-PCR with 58 unpaired arbitrary primers was used to characterize 7 Streptococcus gordonii, 11 Streptococcus sanguis, 2 Streptococcus crista, 5 Streptococcus parasanguis, 18 Streptococcus oralis, and 36 Streptococcus mitis (22 biovar 1 and 14 biovar 2). S. parasanguis 16S rRNA variable region primer RR2 produced species-specific bands with all S. gordonii and S. sanguis. Human V beta 1 T-cell receptor primer 434 yielded concordant genotypic identification of all phenotypically defined S. crista and S. parasanguis, 83% of S. oralis, and 74% of S. mitis biovar 1. Amplicon patterns for S. mitis biovar 2 were heterogeneous. Findings suggest that primers RR2 and 434 in succession will allow rapid identification of genotypic groups corresponding closely to mitis group species established by phenotype.

Identification of mutans and other oral streptococci by random amplified polymorphic DNA analysis

“…It has been shown that the polymorphism in length of the amplified sequences (amplicons) of RAPD fingerprints obtained by AP-PCR can be used to compare bacterial strains [22][23][24], including the groups of mutans streptococci [28,29] and mitis streptococci [30]. In the present study, this application of RAPD analysis to genotyping of oral streptococci confirmed that the RAPD assay can distinguish the species S. mutans and S. sobrinus from each other.…”

“…They include DNA-DNA hybridisation [1,12,14], restriction endonuclease analysis (REA) [16][17][18][19], multilocus enzyme electrophoresis (MLEE) [20], ribotyping, PCR and colony hybridisation with DNA probes [21]. A PCR-based procedure of DNA fingerprinting, called arbitrarily primed PCR (AP-PCR) [22] or randomly amplified polymorphic DNA (RAPD) analysis [23], has been applied to strain identification and typing of oral bacteria [24][25][26][27], oral streptococci [28][29][30] and non-oral streptococci [31][32][33][34]. The present study used RAPD analysis for typing S. mutans and S. sobrinus strains and to explore the potential of this typing method to distinguish these from the other taxa that comprise the group of mutans streptococci.…”

Biological and molecular characterization of Streptococcus crista strains isolated from human dental biofilm by means of arbitrary primers - PCR (AP-PCR)

“…Our results using both primers described by Rudney and Larson (32), and also both primers together with additional primers (OPRs), indicated a varied degree of similarity among the S. crista strains studied herein. The results using the latter method (combination of primers) showed a lower similarity among strains than that obtained when only the RR2 and 434 primers were used.…”

“…The purpose of this work was to biologically characterize different S. crista strains obtained from the dental biofilm of different adult individuals seen at the Dental Clinic of São Francisco University, Bragança Paulista, SP, Brazil by means of molecular techniques, and to compare the results with those published by other groups (3,6,(7)(8)(9)13,18,19,32,44). To the best of our knowledge, no other research group has published in Brazil, any characterization of these types of strains regarding their molecular profile.…”

“…Among the bacterial species present in the dental biofilm, the Streptococcus mitis group (17,32) is considered to be important for its formation. Streptococcus crista is a subspecies belonging to the S. mitis group (10,13,15), and its biochemical characterization is considered to be difficult because of the phenotypic variation exhibited by individuals of the same species (30,31).…”