Arbitrary primed PCR fingerprinting and serotyping of clinical Pseudomonas aeruginosa strains

• Published in: FEMS immunology and medical microbiology Vol. 17; no. 1; pp. 37 - 47
• Main Authors: Hernández, Javier, Ferrús, Marı́a A, Hernández, Manuel, Owen, Robert J
• Format: Journal Article
• Language: English
• Published: Oxford Elsevier B.V 1997; Blackwell
• Subjects: AP-PCR; Bacteriological methods and techniques used in bacteriology; Bacteriology; Biological and medical sciences; Cross Infection - epidemiology; Cross Infection - microbiology; DNA; DNA Fingerprinting; DNA, Bacterial - analysis; Fingerprinting; Fundamental and applied biological sciences. Psychology; Humans; Microbiology; Polymerase chain reaction; Polymerase Chain Reaction - methods; Pseudomonas aeruginosa; Pseudomonas aeruginosa - classification; Pseudomonas aeruginosa - genetics; Pseudomonas aeruginosa - growth & development; Pseudomonas aeruginosa - isolation & purification; Pseudomonas Infections - epidemiology; Pseudomonas Infections - microbiology; Random Amplified Polymorphic DNA Technique; Reproducibility of Results; Serotyping; Polymerase chain reaction; Pseudomonas aeruginosa; AP-PCR; Fingerprinting; DNA; Serotyping; Pseudomonadales; Typing; Random amplified polymorphic DNA; Fingerprint method; Bacteria; Pseudomonadaceae; Genotype; Method; Comparative study
• ISSN: 0928-8244; 1574-695X
• DOI: 10.1016/S0928-8244(96)00103-4

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.