Genotypic Variations of Mutans Streptococci Isolated from Dental Caries by REP-PCR

Main Article Content

Hamzah Abdulrahman Salman
R. Senthilkumar

Abstract

Mutans streptococci (MS) are a group of oral bacteria considered as the main cariogenic organisms. MS consists of several species of genus Streptococcus which are sharing similar phenotypes and genotypes. The aim of this study is to determine the genetic diversity of the core species of clinical strains of Streptococcus mutans, Streptococcus sobrinus and Streptococcus downei by using repitative extragenic palindromic (REP) primer. The DNA of the clinical strains of S. mutans (n=10), S. sobrinus (n=05) and S. downei (n=04) have been employed in the present study, which have been previously isolated from caries active subjects. The DNA of the clinical and reference strains was subjected to PCR amplification using REP primer. The phylogenetic dendrogram is constructed from the REP PCR banding profile by neighbour-joining method using PyElph 1.4 software. The size of the DNA amplicons generated by using REP primer were S. mutans (1500 bp to 250 bp), S. sobrinus (6000 bp to 250 bp) and S. downei (5000 bp to 400 bp). The results present common band at 480 bp in all the clinical strains of S. sobrinus. The current study is the first to demonstrate the genetic variety of S. sobrinus and S. downei by using REP primer. REP-PCR have been found to be a powerful method to study the molecular diversity of S. mutans, S. sobrinus and S. downei. Additionally, further studies are suggested to analyze the species specific bands and also to find the possibility to produce a new specific primer for S. sobrinus.

Downloads

Download data is not yet available.

Article Details

How to Cite
1.
Salman HA, Senthilkumar R. Genotypic Variations of Mutans Streptococci Isolated from Dental Caries by REP-PCR. Baghdad Sci.J [Internet]. 2020Dec.1 [cited 2021Jan.25];17(4):1133. Available from: https://bsj.uobaghdad.edu.iq/index.php/BSJ/article/view/3434
Section
article

References

Billings M, Holtfreter B, Papapanou PN, Mitnik GL, Kocher T, Dye BA. Age‐dependent distribution of periodontitis in two countries: Findings from NHANES 2009 to 2014 and SHIP‐TREND 2008 to 2012. J Clin periodontal. 2018;45:S130-148.

López R, Smith PC, Göstemeyer G, Schwendicke FJ. Ageing, dental caries and periodontal diseases. J Clin Periodonto. 2017;44:S145-S152.

Philip N, Suneja B, Walsh LJ. Ecological approaches to dental caries prevention: paradigm shift or shibboleth?. Caries res. 2018;52:153-65.

Costa S, Martins C, Pinto M, Vasconcelos M, Abreu M. Socioeconomic factors and caries in people between 19 and 60 years of age: an update of a systematic review and meta-analysis of observational studies. Int J Environ Res Public Health. 2018;15:1775.

Frencken JE, Sharma P, Stenhouse L, Green D, Laverty D, Dietrich T. Global epidemiology of dental caries and severe periodontitis–a comprehensive review. J Clin Periodontal. 2017;44:S94-S105.

Bagramian RA, Garcia-Godoy F, Volpe AR. The global increase in dental caries. A pending public health crisis. Am J Dent. 2009;22:3-8.

Baker JL, Edlund A. Exploiting the Oral Microbiome to Prevent Tooth Decay: Has Evolution Already Provided the Best Tools?. Front Microbiol. 2019;9:3323.

Salman HA, Senthilkumar R, Imran K, Selvam KP. Isolation and typing of Streptococcus mutans and Streptococcus sobrinus from caries-active subjects. Contemp Clin Dent. 2017;8:587-93.

Saraithong P, Pattanaporn K, Chen Z, Khongkhunthian S, Laohapensang P, Chhun N, et al. Streptococcus mutans and Streptococcus sobrinus colonization and caries experience in 3-and 5-year-old Thai children. Clin Oral Investig. 2015;19:1955-64.

Oda Y, Hayashi F, Okada MJBOH. Longitudinal study of dental caries incidence associated with Streptococcus mutans and Streptococcus sobrinus in patients with intellectual disabilities. BMC Oral Health. 2015;2;15:102.

Salman HA, Senthilkumar R, Mahmood BS, Imran K. Detection and characterization of Streptococcus downei, a rare bacterial species of mutans streptococci from caries active patients. Indian J Dent Res. 2019;30:579-82.

Cheon K, Moser SA, Wiener HW, Whiddon J, Momeni SS, Ruby JD, et al. Characteristics of Streptococcus mutans genotypes and dental caries in children. Eur J Oral Sci. 2013;121148-155.

Braga MP, Piovesan A, Valarini N, Maciel SM, Andrade FBd, Poli-Frederico RC. Genotypic diversity and virulence factors of Streptococcus mutans in caries-free and caries-active individuals. Braz Arch Biol Technol. 2013;56:241-8.

Bedoya-Correa CM, Rodríguez RJ, Parada-Sanchez MT. Genomic and phenotypic diversity of Streptococcus mutans. J Oral Biosci. 2019;61:22-31.

Mineyama R, Yoshino S, Maeda NJ. DNA fingerprinting of isolates of Streptococcus mutans by pulsed-field gel electrophoresis. Microbiol Research. 2007;162:244-9.

Valdez RMA, Duque C, Caiaffa KS, dos Santos VR, de Aguiar Loesch ML, Colombo NH, et al. Genotypic diversity and phenotypic traits of Streptococcus mutans isolates and their relation to severity of early childhood caries. BMC Oral Health. 2017:17:115.

Momeni SS, Whiddon J, Moser SA, Cheon K, Ruby JD, Childers NK. Comparative genotyping of Streptococcus mutans by repetitive extragenic palindromic polymerase chain reaction and multilocus sequence typing. Mol Oral Microbiol. 2013;2818-27.

Moser SA, Mitchell SC, Ruby JD, Momeni S, Osgood RC, Whiddon J, et al. Repetitive extragenic palindromic PCR for study of Streptococcus mutans diversity and transmission in human populations. J Clin Microbiol. 2010:48599-602

Versalovic J, Koeuth T, Lupski RJ. Distribution of repetitive DNA sequences in eubacteria and application to finerpriting of bacterial enomes. Nucleic Acids Res. 1991;19:6823-31.

Fakruddin M, Mannan B, Shahnewaj K, Mazumdar RM, Chowdhury A, Hossain N. Identification and characterization of microorganisms: DNA-fingerprinting methods. Songklanakarin J Scien Technol. 2013;35:397-404.

Stern MJ, Ames GF-L, Smith NH, Robinson EC, Higgins CF. Repetitive extragenic palindromic sequences: a major component of the bacterial genome. Cell. 1984;37:1015-26.

Healy M, Huong J, Bittner T, Lising M, Frye S, Raza S, et al. Microbial DNA typing by automated repetitive-sequence-based PCR. J Clin Microbiol. 2005;43199-207.

Momeni SS, Whiddon J, Cheon K, Ghazal T, Moser SA, Childers NK. Genetic Diversity and Evidence for Transmission of Streptococcus mutans by DiversiLab rep-PCR. J Microbiol Methods. 2016;128108-117.

Salman HA, Kumar RS, Babu NC, Imran K. First detection and characterization of Streptococcus dentapri from caries active subject. J Clin Diagn Res. 2017;11:DM01 3.

Versalovic J, Schneider M, De Bruijn FJ, Lupski JR. Genomic fingerprinting of bacteria using repetitive sequence-based polymerase chain reaction. Methods Mol Cell Biol. 1994;5:25-40.

Pavel AB, Vasile CI. PyElph - a software tool for gel images analysis and phylogenetics. BMC Bioinformatics. 2012;13:9.

Okada T, Takada K, Fujita K, Ikemi T, Osgood RC, Childers NK, et al. Differentiation of banding patterns between Streptococcus mutans and Streptococcus sobrinus isolates in rep-PCR using ERIC primer. J Oral Microbiol. 2011;3:7190.

Alam S, Brailsford SR, Whiley RA, Beighton D. PCR-based methods for genotyping viridans group streptococci. J Clin Microbiol. 1999; 37: 2772_6.

Clarridge IIIJ, Osting C, Jalali M, Osborne J, Waddington M. Genotypic and phenotypic characterization of “Streptococcus milleri” group isolates from a veterans administration hospital population. J Clin Microbiol. 1999;37:3681-7.

Cheon K, Moser SA, Whiddon J, Osgood RC, Momeni S, Ruby JD, et al. Genetic diversity of plaque mutans streptococci with rep-PCR. J Dent Res. 2011; 90331-35.

Whiley RA, Russell RR, Hardie J, Beighton D. Streptococcus downei sp. nov. for strains previously described as Streptococcus mutans serotype h. Int J Syst Bacteriol. 1988;38:25 9.