Molecular epidemiology of Pseudomonas aeruginosa from cystic fibrosis in Sicily: genome macrorestriction analysis and rapid PCR-ribotyping.
This study addresses the epidemiologic relatedness of a collection of Pseudomonas aeruginosa isolates from cystic fibrosis patients attending the Pediatric Clinic, Catania, Sicily. Genome macrorestriction analysis after pulsed field gel electrophoresis (PFGE) was used to characterise all strains. Furthermore, a rapid typing procedure, developed in this study, based on polymerase chain reaction amplified ribosomal DNA spacer polymorphisms (PCR-ribotyping), straight from bacterial cultures, was used. On the basis of macrorestriction analysis after PFGE, persistence of infection was shown in all patients; two cross-transmission episodes were identified in the nosocomial as well as in the familiar environment. PCR-ribotyping proved to be useful for a DNA-based identification test, suitable for screening purposes. The rapid amplification protocol here tested is proposed to evaluate the discriminatory power of other specific target sequences in PCR-based typing assays, for epidemiologic purposes.
Agodi
A
A
Institute of General Biology, University of Catania, Italy.
Sciacca
A
A
Campanile
F
F
Messina
C
C
Barchitta
M
M
Sciacca
S
S
Stefani
S
S
D000293:Adolescent; D000900:Anti-Bacterial Agents / Q000494:pharmacology; D002648:Child; D002675:Child, Preschool; D003550:Cystic Fibrosis / Q000150:complications / Q000382:microbiology*; D004269:DNA, Bacterial / Q000235:genetics / Q000378:metabolism; D021901:DNA, Intergenic / Q000235:genetics / Q000378:metabolism; D004275:DNA, Ribosomal / Q000235:genetics / Q000378:metabolism; D015252:Deoxyribonucleases, Type II Site-Specific / Q000378:metabolism; D004352:Drug Resistance, Microbial; D005260:Female; D016680:Genome, Bacterial; D005838:Genotype; D006801:Humans; D007223:Infant; D008297:Male; D008826:Microbial Sensitivity Tests; D016133:Polymerase Chain Reaction; D011110:Polymorphism, Genetic; D011552:Pseudomonas Infections / Q000150:complications / Q000453:epidemiology* / Q000382:microbiology / Q000635:transmission; D011550:Pseudomonas aeruginosa / Q000145:classification / Q000187:drug effects / Q000235:genetics*; D015183:Restriction Mapping; D021521:Ribotyping; D012802:Sicily
D000900:Anti-Bacterial Agents; D004269:DNA, Bacterial; D021901:DNA, Intergenic; D004275:DNA, Ribosomal; C024669:endodeoxyribonuclease AluI; D015252:Deoxyribonucleases, Type II Site-Specific