title: Evaluation and comparison of random amplification of polymorphic DNA, pulsed-field gel electrophoresis and ADSRRS-fingerprinting for typing Serratia marcescens outbreaks.
abstract: Amplification of DNA fragments surrounding rare restriction sites (ADSRRS-fingerprinting) is a novel assay based on suppression of polymerase chain reaction (PCR). This phenomenon allows the amplification of only a limited subset of DNA fragments, since only those with two different oligonucleotides ligated at the ends of complementary DNA strands are amplified in the PCR. The DNA fragments can be easily analyzed on polyacrylamide gels, stained with ethidium bromide. We have implemented this method using a set of clinical Serratia marcescens isolates from three outbreaks ongoing in the Public Hospital in Gdańsk (Poland). Clustering of ADSRRS-fingerprinting data matched epidemiological, microbiological, random amplification of polymorphic DNA (RAPD) and pulsed-field gel electrophoresis (PFGE) data. Based on this study, we found that there is at least a similar power of discrimination between the present 'gold-standard' PFGE and the novel method, ADSRRS-fingerprinting. Although the ADSRRS-fingerprinting method may appear to be more complex than the RAPD technique, we found it fast and reproducible.
authors.affiliation: Department of Microbiology, Technical University of Gdańsk, ul G Narutowicza 11/12, 80-952 Gdańsk, Poland.
authors.forename: Beata
authors.initials: B
authors.lastname: Krawczyk
authors.forename: Lukasz
authors.initials: L
authors.lastname: Naumiuk
authors.forename: Krzysztof
authors.initials: K
authors.lastname: Lewandowski
authors.forename: Anna
authors.initials: A
authors.lastname: Baraniak
authors.forename: Marek
authors.initials: M
authors.lastname: Gniadkowski
authors.forename: Alfred
authors.initials: A
authors.lastname: Samet
authors.forename: Józef
authors.initials: J
authors.lastname: Kur
mesh_terms: D000900:Anti-Bacterial Agents / Q000494:pharmacology; D015373:Bacterial Typing Techniques / Q000379:methods*; D002876:Chromosomes, Bacterial / Q000737:chemistry; D016172:DNA Fingerprinting / Q000379:methods*; D004269:DNA, Bacterial / Q000737:chemistry / Q000302:isolation & purification; D004196:Disease Outbreaks; D024881:Drug Resistance, Bacterial; D016521:Electrophoresis, Gel, Pulsed-Field; D004591:Electrophoresis, Polyacrylamide Gel; D005838:Genotype; D006801:Humans; D008826:Microbial Sensitivity Tests; D021141:Nucleic Acid Amplification Techniques / Q000379:methods; D016133:Polymerase Chain Reaction / Q000379:methods; D012150:Polymorphism, Restriction Fragment Length; D019105:Random Amplified Polymorphic DNA Technique; D015203:Reproducibility of Results; D016868:Serratia Infections / Q000453:epidemiology* / Q000382:microbiology*; D012706:Serratia marcescens / Q000145:classification* / Q000235:genetics / Q000302:isolation & purification
chemical_list: D000900:Anti-Bacterial Agents; D004269:DNA, Bacterial