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Arbitrary primed PCR fingerprinting and serotyping of clinical Pseudomonas aeruginosa strains
Authors:Javier Herná  ndez ,Marí  a A Ferrú  s,Manuel Herná  ndez,Robert J Owen
Affiliation:Departamento de Biotecnología, Universidad Politécnica, Camino de Vera 14, 46022 Valencia, Spain;National Collection of Type Cultures, Central Public Health Laboratory, 61 Colindale Avenue, London NW9 5HT, UK
Abstract:Arbitrary primed PCR (AP-PCR) analysis was compared with serotyping as a means of high-resolution typing of Pseudomonas aeruginosa. Seventy-four isolates from 3 different hospitals and 18 reference strains were studied. Serotyping provided good index of discrimination, although eleven isolates could not be serotyped. Genomic DNA was amplified with a single 10 nucleotide primer (sequence 5′-AGG GGT CTT G-3′). The strains were genetically diverse and 61 different AP-PCR profiles of 2–7 bands between 0.3 and 2.4 kb were obtained. AP-PCR profiles were not consistently associated with serotypes, but they clearly subtyped strains of the same serotype. Numerical analysis of AP-PCR patterns defined 7 groups at the 55% similarity level, and identified predominant strains in each hospital. The results show that AP-PCR analysis provides a simple and practical approach to typing P. aeruginosa that is more discriminatory than traditional serotyping scheme. We suggest that maximum discrimination can be achieved by a combination of both methods.
Keywords:Pseudomonas aeruginosa    DNA    Polymerase chain reaction    AP-PCR    Serotyping    Fingerprinting
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