Optimisation of AP-PCR fingerprinting discriminatory power for clinical isolates of Pseudomonas aeruginosa.
Recently methods based on analysis of arbitrarily amplified target sites of microorganism genomes have been extensively applied in microbiological studies. The range of their applications is limited by problems with discrimination and reproducibility resulting from lack of standardised and reliable methods of optimisation. By orthogonal-array optimisation most advantageous and optimal parameters for highly discriminatory primers (CagA2+CMVin2) were selected and efficient AP-PCR (arbitrarily primed-polymerase chain reaction) fingerprinting conditions for Pseudomonas aeruginosa isolates were set up. Stable and multiplex amplicon profiles obtained in this study revealed high level of intraspecies DNA polymorphism among 20 analysed clinical strains of P. aeruginosa proving optimised AP-PCR fingerprinting to be useful in epidemiological typing of the species.
Dabrowski
Waldemar
W
Department of Food Microbiology, Faculty of Food Sciences and Fisheries, Agricultural University of Szczecin, Papieza Pawla VI 3, 71-459, Szczecin, Poland. wm.dabrowski@tz.ar.szczecin.pl
Czekajlo-Kolodziej
Urszula
U
Medrala
Dagmara
D
Giedrys-Kalemba
Stefania
S
D016172:DNA Fingerprinting / Q000379:methods*; D017931:DNA Primers; D004269:DNA, Bacterial / Q000032:analysis; D016680:Genome, Bacterial; D016133:Polymerase Chain Reaction / Q000379:methods; D011550:Pseudomonas aeruginosa / Q000235:genetics* / Q000302:isolation & purification*
D017931:DNA Primers; D004269:DNA, Bacterial