Open AccessOptimisation of AP–PCR fingerprinting discriminatory power for clinical isolates of Pseudomonas aeruginosa
Author(s) -
Da̧browski Waldemar,
CzekajłoKołodziej Urszula,
Mȩdrala Dagmara,
GiedrysKalemba Stefania
Publication year - 2003
Publication title -
fems microbiology letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.899
H-Index - 151
eISSN - 1574-6968
pISSN - 0378-1097
DOI - 10.1016/s0378-1097(02)01183-7
Subject(s) - amplicon , pseudomonas aeruginosa , dna profiling , biology , multiplex polymerase chain reaction , polymerase chain reaction , typing , microbiology and biotechnology , genetics , computational biology , dna , bacteria , gene
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.