title: Rapid genotyping of Achromobacter xylosoxidans, Acinetobacter baumannii, Klebsiella pneumoniae, Pseudomonas aeruginosa and Stenotrophomonas maltophilia isolates using melting curve analysis of RAPD-generated DNA fragments (McRAPD).
abstract: Typing of bacteria is important for monitoring newly emerging pathogens and for examining local outbreaks. We evaluated the randomly amplified polymorphic DNA technique in combination with melting curve analysis (McRAPD) of the amplified DNA fragments to genotype isolates from five Gram-negative species, i.e. Achromobacter xylosoxidans, Acinetobacter baumannii, Klebsiella pneumoniae, Pseudomonas aeruginosa and Stenotrophomonas maltophilia. By determining the melting temperature peaks of the amplified DNA fragments, we were able to distinguish the different genotypes of isolates, as they had been assessed by other genotyping techniques, i.e. agarose gel electrophoresis of RAPD fragments, multilocus sequence typing and/or AFLP™. According to our results, McRAPD may offer the possibility of genotyping a limited number of bacterial isolates, e.g. in case of suspicion of hospital outbreak, via a less costly, more rapid, less laborious and more user-friendly technique than RAPD followed by electrophoresis.
authors.affiliation: Laboratory for Bacteriology Research (LBR), Ghent University Hospital, University of Ghent, De Pintelaan 185, 9000 Ghent, Belgium. Deschaght.Pieter@UGent.be
authors.forename: Pieter
authors.initials: P
authors.lastname: Deschaght
authors.forename: Leen
authors.initials: L
authors.lastname: Van Simaey
authors.forename: Ellen
authors.initials: E
authors.lastname: Decat
authors.forename: Els
authors.initials: E
authors.lastname: Van Mechelen
authors.forename: Sylvain
authors.initials: S
authors.lastname: Brisse
authors.forename: Mario
authors.initials: M
authors.lastname: Vaneechoutte
mesh_terms: D042441:Achromobacter denitrificans / Q000145:classification / Q000235:genetics / Q000302:isolation & purification*; D040981:Acinetobacter baumannii / Q000145:classification / Q000235:genetics / Q000302:isolation & purification*; D015373:Bacterial Typing Techniques / Q000379:methods*; D004269:DNA, Bacterial / Q000235:genetics; D005838:Genotype; D016905:Gram-Negative Bacterial Infections / Q000382:microbiology*; D006801:Humans; D007711:Klebsiella pneumoniae / Q000145:classification / Q000235:genetics / Q000302:isolation & purification*; D011550:Pseudomonas aeruginosa / Q000145:classification / Q000235:genetics / Q000302:isolation & purification*; D019105:Random Amplified Polymorphic DNA Technique / Q000379:methods*; D020615:Stenotrophomonas maltophilia / Q000145:classification / Q000235:genetics / Q000302:isolation & purification*; D044366:Transition Temperature
chemical_list: D004269:DNA, Bacterial