Enterococcus faecalis patterning by pulsed-field gradient gel electrophoresis and polymerase chain reaction

Abstract

Pulsed field gel electrophoresis ( PFGE ) using Sma I resctriction enzyme and polymerase chain reaction ( PCR ) employing REP-1 ( 5’ IIIICGICGICATCIGGC 3’ ), REP-2R (5’ ICGICTTATCIGGCCTAC 3’ ) and JB1-(5’GATTTTATGGCCGTCCGC3’) primers were used to study genomic DNA from 10 Enterococcus faecium clinical samples. By PFGE we found 10 resctriction patterns varying from 10 to 20 fragments. Using REP-1/2R primers we found 7 banding patterns, changind from 4 to 10 fragments, while with JB1-PCR primer it was found 6 banding patterns changing from 4 to 6 fragments. Identical genotypes were found by both REP-PCR and JB1-PCR however only 2 strains were considered identical by two PCR techniques simultaneosly. Due to low number of isolates it was not applied any statistical method to analyse the prevalence of genotypes among the samples. Genetic heterogeneity was observed among isolates by both PFGE and PCR techniques and the restriction pattern was easier interpreted than PCR banding pattern. The primer JB1 5’GATTTTATGGCCGTCCGC3’ was used by the first time to study Enterococcus faecalis and was a useful tool to discriminate strains of this specie