Enhanced resolution of random amplified polymorphic DNA genotyping of Pseudomonas aeruginosa.
OBJECTIVE
To develop a rapid, sensitive and reproducible screening test for the detection of nosocomial spreading of Pseudomonas aeruginosa.


RESULTS
Ps. aeruginosa genomic DNA extraction, RAPD-PCR, electrophoresis on acrylamide gel and silver staining were performed by using standardized reagents and conditions. The results were compared with the agarose gel electrophoresis followed by ethidium bromide staining.


CONCLUSIONS
The coupling of acrylamide gel electrophoresis and silver staining gave about 80% more DNA bands than the traditional method, allowing a finer discrimination among different Ps. aeruginosa strains.


CONCLUSIONS
By enhancing the resolution of the electrophoretic separation and the sensitivity of the staining, random amplification could be easily applied to the surveillance and prevention of nosocomial infections by clinical microbiology laboratories.
Dipartimento di Patologia Sperimentale, Biotecnologie Mediche, Infettivologia ed Epidemiologia, Unità Operativa di Microbiologia Universitaria, Azienda Ospedaliera Universitaria Pisana, Pisa, Italy. s.barnini@ao-pisa.toscana.it
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D015373:Bacterial Typing Techniques; D003428:Cross Infection / Q000382:microbiology; D004269:DNA, Bacterial / Q000032:analysis; D004591:Electrophoresis, Polyacrylamide Gel; D005838:Genotype; D006801:Humans; D011552:Pseudomonas Infections / Q000382:microbiology; D011550:Pseudomonas aeruginosa / Q000145:classification* / Q000235:genetics*; D019105:Random Amplified Polymorphic DNA Technique / Q000379:methods*; D016622:Silver Staining / Q000379:methods; D013997:Time Factors
D004269:DNA, Bacterial