Evaluation of (GTG)5-PCR for rapid identification of Streptococcus mutans

Abstract

Repetitive sequence-based polymerase chain reaction (PCR) fingerprinting using the (GTG)(5) primer was applied for fast screening of bacterial strains isolated from dental plaque of early childhood caries (ECC)-affected children. A group of 29 Gram-positive bacteria was separated into a homogeneous cluster together with Streptococcus mutans reference strains and constituted an aberrant branch after the numerical analysis of (GTG)(5)-PCR fingerprints. Automated ribotyping with EcoRI restriction enzyme (RiboPrinter microbial characterization system) revealed high genetic heterogeneity among the tested group and proved to be a good tool for strain-typing purposes. Further characterization of the studied strains was achieved by extensive phenotyping and whole-cell protein fingerprinting and confirmed all the strains as S. mutans representatives. Obtained results showed rep-PCR fingerprinting with the (GTG)(5) primer to be a fast and reliable method for identification of S. mutans.

Antibiotic resistance in faecal bacteria (Escherichia coli, Enterococcus spp.) in feral pigeons

“…(2008). Briefly, the total genomic DNA isolated by alkaline extraction procedure was amplified using the (GTG) 5 primer (5′‐GTG GTG GTG GTG GTG‐3′) with the PCR programme and conditions described previously (Svec et al. 2008).…”

“…The (GTG) 5 -PCR fingerprinting method was performed according to Svec et al (2008). Briefly, the total genomic DNA isolated by alkaline extraction procedure was amplified using the (GTG) 5 primer (5¢-GTG GTG GTG GTG GTG-3¢) with the PCR programme and conditions described previously (Svec et al 2008). Obtained PCR products were separated for 16 h at 55 V in 1AE5% (w ⁄ v) agarose gels (20 x 25 cm).…”

TRS-based PCR as a potential tool for inter-serovar discrimination of Salmonella Enteritidis, S. Typhimurium, S. Infantis, S. Virchow, S. Hadar, S. Newport and S. Anatum

“…In other studies, methods using the (GTG) 5 primer were evaluated for the identification of Streptococcus mutans , Bacillus spp. and Klebsiella isolates [ 21 – 23 ]. However, these studies lacked reproducibility analyses, and there were some inaccuracies in the grouping of the bacterial isolates.…”

Comparison of (GTG)5-oligonucleotide and ribosomal intergenic transcribed spacer (ITS)-PCR for molecular typing of Klebsiella isolates

“…In some recent studies, (GTG) 5 -PCR fingerprint analysis has been used for molecular typing of Acinetobacter baumanii (Huys, et al, 2005), Salmonella enterica (Rasschaert et al, 2005), Campylobacter concisus (Matsheka et al, 2006), Enterococcus faecium (Svec et al, 2005;Jurkovic et al, 2007), Escherichia coli (Mohapatra et al, 2007: Mohapatra et al, 2008, Streptococcus mutans (Svec et al, 2008), and for the identification of lactic acid bacteria isolated from human blood cultures (Svec et al, 2007). However, so far no data are available demonstrating the potential use of (GTG) 5 -fingerprint analysis as a molecular means to differentiate Klebsiella spp.…”