Journal Article

Evaluation and comparison of random amplification of polymorphic DNA, pulsed-field gel electrophoresis and ADSRRS-fingerprinting for typing Serratia marcescens outbreaks

Beata Krawczyk, Łukasz Naumiuk, Krzysztof Lewandowski, Anna Baraniak, Marek Gniadkowski, Alfred Samet and Józef Kur

in FEMS Immunology & Medical Microbiology

Published on behalf of Federation of Microbiological Societies

Volume 38, issue 3, pages 241-248
Published in print October 2003 | ISSN: 0928-8244
Published online January 2006 | e-ISSN: 1574-695X | DOI: https://dx.doi.org/10.1016/S0928-8244(03)00149-4
Evaluation and comparison of random amplification of polymorphic DNA, pulsed-field gel electrophoresis and ADSRRS-fingerprinting for typing Serratia marcescens outbreaks

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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.

Keywords: Enterobacteriaceae; PCR; PCR suppression; PCR-fingerprinting; Random amplification of polymorphic DNA; Pulsed-field gel electrophoresis; ADSRRS-fingerprinting

Journal Article.  4326 words.  Illustrated.

Subjects: Medical Microbiology and Virology ; Biotechnology ; Genetics and Genomics ; Microbiology ; Molecular and Cell Biology

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