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Advisory Committee Chair

Noel K Childers

Advisory Committee Members

Susan Hollingshead

Stephen A Moser

John D Ruby

Document Type

Thesis

Date of Award

2010

Degree Name by School

Master of Dentistry (MDent) School of Dentistry

Abstract

ABSTRACT The role of Streptococcus mutans has been extensively studied using a variety of genotyping techniques. Repetitive extragenic palindromic polymerase chain reaction (rep-PCR) is an effective tool for screening large-scale epidemiological studies with high discriminatory power and reproducibility. For this study, multilocus sequence typing (MLST) analysis is used to evaluate genotypes previously identified as unique using rep-PCR. Twenty-two unique S. mutans rep- PCR genotypes were selected from a longitudinal study. Four additional isolates were selected from each genotype group of the 6 most commonly occurring genotypes (n=30) for further analysis. Real-time PCR was performed using eight housekeeping S. mutans gene loci and products sequenced. Sequence data analysis was performed using CLC DNA Workbench with MLST module. Concatenated sequences (3366 bp) were evaluated with MEGA using minimum evolution method with bootstrap. All 22 rep-PCR genotypes were unique by MLST analysis. For rep-PCR genotype groups, MLST groups matched rep-PCR for 3/6 (50%); three groups were further distinguished with MLST analysis. Rep-PCR results were consistent with those found using MLST, although MLST can provide further discrimination for some isolates. Rep-PCR is an effective tool for screening large-scale populations studies of S. mutans for subsequent MLST analysis.

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