Comparisons of Two Proteomic Analyses of Non-Mucoid and Mucoid Pseudomonas aeruginosa Clinical Isolates from a Cystic Fibrosis Patient (original) (raw)
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Microbiology (Reading, England), 2000
Isolates of Pseudomonas aeruginosa from chronic lung infections in cystic fibrosis (CF) patients have phenotypes distinct from those initially infecting CF patients, as well as from other clinical or environmental isolates. To gain a better understanding of the differences in these isolates, protein expression was followed using two-dimensional (2-D) gel electrophoresis and protein identification by peptide sequencing using micro-capillary column liquid chromatography-tandem mass spectrometry (microLC/MS/MS). The isolates selected for this analysis were from the sputum of a CF patient: strain 383 had a nonmucoid phenotype typical of isolates from the environment, and strain 2192, obtained from the same patient, had a mucoid phenotype typical of isolates from chronic CF lung infections. Strains 383 and 2192 were confirmed to be genetically identical by restriction endonuclease analysis, random amplified polymorphic DNA-PCR, and pulsed-field gel electrophoresis. Conditions of protein ...
Clinical Microbiology and Infection, 2011
Background: Chronic lung infection with the bacterium Pseudomonas aeruginosa is one of the hallmarks of cystic fibrosis (CF) and is associated with worsening lung function, increased hospitalisation and reduced life expectancy. A virulent clonal strain of P. aeruginosa (Australian epidemic strain I; AES-I) has been found to be widespread in CF patients in eastern Australia. Methods: Suppression subtractive hybridization (SSH) was employed to identify genetic sequences that are present in the AES-I strain but absent from the sequenced reference strain PAO1. We used PCR to evaluate the distribution of several of the AES-I loci amongst a collection of 188 P. aeruginosa isolates which was comprised of 35 AES-I isolates (as determined by PFGE), 78 non-AES-I CF isolates including other epidemic CF strains as well as 69 P. aeruginosa isolates from other clinical and environmental sources. We have identified a unique AES-I genetic locus that is present in all 35 AES-I isolates tested and not present in any of the other 153 P. aeruginosa strains examined. We have used this unique AES-I locus to develop a diagnostic PCR and a real-time PCR assay to detect the presence of P. aeruginosa and AES-I in patient sputum samples. We have developed diagnostic PCR assays that are 100% sensitive and 100% specific for the P. aeruginosa strain AES-I. We have also shown that Whatman FTA ® Elute cards may be used with PCR-based assays to rapidly detect the presence of P. aeruginosa strains in CF sputum.
Brazilian Journal of Infectious Diseases, 2010
The present study had as objective to evaluate the genotypic diversity and biological characteristics, such as hemolysin, protease, elastase of 56 clinical strains of Pseudomonas aeruginosa isolated from 13 cystic fi brosis (CF) patients attending at the School Hospital of Campinas State University (UNICAMP), Brazil. Genotypic diversity has been determined by Ribotyping (RT) and the pattern of the enterobacterial repetitive intergenic consensus PCR (ERIC-PCR) of each strain. The production of elastase was signifi cantly different only among mucoid and nonmucoid isolates. Joint results obtained by (RT) and ERIC-PCR methods were able to discriminate all strains isolated from both the same and different patients. Additionally, we observed four strain clusters with low diversity. The most infective strains were located in just two clusters. These results suggest that either there is a strong selection towards a specifi c genotype or that specifi c isolates could be responsible for the initial and subsequent colonization processes. More studies are necessary to know if these conclusions can be generalized for the general CF population.
Epidemiologic characterization of Pseudomonas aeruginosa in patients with cystic fibrosis
Clinical Microbiology and Infection, 2000
Objective To determine persistence and variability of colonization with Pseudomonas aeruginosa in cystic ¢brosis patients over long time periods, and to look for possible cross-colonization. Methods In total, 469 Pseudomonas aeruginosa isolates were obtained from 30 patients during the period from April 1994 to April 1996.The sources were mainly sputum and a few deep throat swabs. All grown strains dissimilar in macromorphology were processed separately.Typing with PFGE was carried out by contour-clamped homogeneous electric ¢eld electrophoresis. Genomic DNAwas subjected to the rarecutting restriction enzyme SpeI. For pyocin typing, the procedure described by Fyfe was applied. Results After typing with PFGE, we observed 40 restriction pro¢les. Eighteen di¡erent pyocin types were found.The most frequent pyocin type was type 3, followed by types 1 and 5.Twenty-two patients were persistently colonized by one clone speci¢c and di¡erent for each patient, and four were co-colonized by a second clone also di¡erent for each of these patients. Cross-colonization had apparently been rare in the cystic ¢brosis center of Leipzig. Conclusions Typing with PFGE is well suited for detailed investigations of colonization with Pseudomonas aeruginosa in cystic ¢brosis patients. Pyocin typing can provide additional information for epidemiologic purposes.