ABSTRACT
Forty-seven Enterococcus faecium strains from different sources were evaluated by restriction endonuclease analysis (REA) of total chromosomal DNA. Strains from chicken, pork, and humans were clearly divided into separate clusters, whereas strains from different countries, strains with different antibiotic resistance profiles, or clinical and healthy-subject strains were not.
Subject(s)
Enterococcus faecium/classification , Enterococcus faecium/genetics , Gram-Positive Bacterial Infections/microbiology , Restriction Mapping , Animals , Anti-Bacterial Agents/pharmacology , Deoxyribonuclease EcoRI/metabolism , Deoxyribonucleases, Type II Site-Specific/metabolism , Drug Resistance, Microbial , Enterococcus faecium/drug effects , Humans , Phylogeny , ProhibitinsABSTRACT
Randomly amplified polymorphic DNA (RAPD) has been used for rapid typing of Lactobacillus plantarum strains. RAPD was used with either purified chromosomal DNA serving as template in the polymerase chain reaction, or with crude cell extracts, and using a 9-mer primer with 80% G+C content. Amplified DNA was visualized by ethidium bromide staining after separation on agarose gels. Patterns from 20 Lact. plantarum strains and two Lact. pentosus strains were analysed using the Pearson products moment correlation coefficient (r) and the unweighted pair group method with arithmetic averages (UPGMA). With some exceptions, the two sources of template DNA gave the same clusters and subclusters of strains at the similarity level of 50%. About 50% of the strains could be individually separated from all the other tested strains. The buffer brand, the amount of primer and crude cell extract used in the PCR-step were crucial for the final pattern.
Subject(s)
Bacterial Typing Techniques , Lactobacillus/classification , Random Amplified Polymorphic DNA Technique , Base Composition , Base Sequence , Cluster Analysis , DNA Primers/genetics , DNA, Bacterial/genetics , DNA, Bacterial/isolation & purification , Lactobacillus/genetics , Molecular Sequence DataABSTRACT
The use of randomly amplified polymorphic DNA (RAPD) for rapid, reliable, and easily interpreted identification of enterococci was evaluated. Nineteen type strains of Enterococcus, 12 reference strains, and 114 clinical isolates of Enterococcus were analyzed. Discrimination was obtained between most type strains, the exceptions being Ent. casseliflavus and Ent. flavescens, which had relatively similar RAPD-profiles. Ent. faecalis and Ent. faecium were readily separated, and Ent. gallinarum and Ent. durans could also be identified. Extracts to be used in the polymerase chain reaction (PCR) were prepared directly from agar plate colonies, which made it possible to complete the identification procedure in one day. RAPD was proved to be a fast and reliable method for identification of most Enterococcus spp. of clinical significance.
Subject(s)
Enterococcus/isolation & purification , Random Amplified Polymorphic DNA Technique , Enterococcus/geneticsABSTRACT
Two hundred and seventy-seven Enterococcus isolates representing the predominating enterococcal flora of retailed chicken and pork were identified by phenotypical features, and by random amplified polymorphic DNA. The resistance to nine different antibiotics was determined. Enterococcus faecium was the most frequently occurring species in both Swedish and Danish chicken. Enterococcus faecalis and unidentified groups of Enterococcus dominated in pork. Seventy-three per cent of the Enterococcus isolates from Swedish chicken were resistant to one or more of the tested antibiotics. The corresponding values for Swedish pork, Danish chicken and Danish pork were 9%, 55% and 14%, respectively. Tetracycline resistance was most frequent in isolates from Danish pork and Swedish chicken, while erythromycin resistance was most frequent in Danish chicken. Strains with acquired vancomycin resistance, mainly Ent. faecium, were found only on Danish chicken, except for one isolate from Danish pork. All vancomycin-resistant isolates contained the vanA gene. Vancomycin resistance could be transferred to a vancomycin-sensitive Ent. faecium strain from four of nine tested donor strains.