title: Amplification of a single-locus variable-number direct repeats with restriction fragment length polymorphism (DR-PCR/RFLP) for genetic typing of Acinetobacter baumannii strains.
abstract: In search of an effective DNA typing technique for Acinetobacter baumannii strains for hospital epidemiology use, the performance and convenience of a new target sequence was evaluated. Using known genomic sequences of Acinetobacter baumannii strains AR 319754 and ATCC 17978, we developed single-locus variable-number direct-repeat analysis using polymerase chain reaction-restriction fragment length polymorphism (DR-PCR/RFLP) method. A total of 90 Acinetobacter baumannii strains isolated from patients of the Clinical Hospital in Bydgoszcz, Poland, were examined. Initially, all strains were typed using macrorestriction analysis of the chromosomal DNA by pulsed-field gel electrophoresis (REA-PFGE). Digestion of the chromosomal DNA with the ApaI endonuclease and separation of the fragments by PFGE revealed 21 unique types. Application of DR-PCR/RFLP resulted in recognition of 12 clusters. The results showed that the DR-PCR/RFLP method is less discriminatory than REA-PFGE, however, the novel genotyping method can be used as an alternative technique for generating DNA profiles in epidemiological studies of intra-species genetic relatedness of Acinetobacter baumannii strains.
authors.affiliation: Department of Biology and Ecology, Academy of Physical Education and Sport, Gdańsk, Poland. azaleska@awf.gda.pl
authors.forename: Alicja
authors.initials: A
authors.lastname: Nowak-Zaleska
authors.forename: Beata
authors.initials: B
authors.lastname: Krawczyk
authors.forename: Roman
authors.initials: R
authors.lastname: Kotłowski
authors.forename: Agnieszka
authors.initials: A
authors.lastname: Mikucka
authors.forename: Eugenia
authors.initials: E
authors.lastname: Gospodarek
mesh_terms: D040981:Acinetobacter baumannii / Q000145:classification / Q000235:genetics*; D015373:Bacterial Typing Techniques / Q000379:methods*; D001483:Base Sequence; D002876:Chromosomes, Bacterial; D004269:DNA, Bacterial / Q000032:analysis; D016521:Electrophoresis, Gel, Pulsed-Field; D005838:Genotype; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction; D012150:Polymorphism, Restriction Fragment Length; D012091:Repetitive Sequences, Nucleic Acid; D017422:Sequence Analysis, DNA
chemical_list: D004269:DNA, Bacterial