Evaluation and comparison of random amplification of polymorphic DNA, pulsed-field gel electrophoresis and ADSRRS-fingerprinting for typing Serratia marcescens outbreaks.
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.
Krawczyk
Beata
B
Department of Microbiology, Technical University of Gdańsk, ul G Narutowicza 11/12, 80-952 Gdańsk, Poland.
Naumiuk
Lukasz
L
Lewandowski
Krzysztof
K
Baraniak
Anna
A
Gniadkowski
Marek
M
Samet
Alfred
A
Kur
Józef
J
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
D000900:Anti-Bacterial Agents; D004269:DNA, Bacterial