RESUMO
INTRODUCTION: Pseudomonas aeruginosa is able to colonize the lungs of cystic fibrosis patients (CF) in an adaptive process that results in the selection of a dominant strain through a process of genetic variation. METHODS: One hundred and twenty tree isolates of P. aeruginosa were sequentially recovered from 6 CF patients during the routine follow-up or exacerbations over periods of 2 to 12 years in the Ramon y Cajal University Hospital (Madrid, Spain). Another 13 isolates were obtained from a single CF patient in a short-term study. They were analysed by restriction fragment length polymorphism (RFLP) and sequencing of mucA and fpvA genes, which code for the alginate biosynthesis regulator and a pyoverdin receptor, respectively, and their antibiotic susceptibility was studied by microdilution. RESULTS: A dominant colonising strain was found in each patient based on the RFLP profile. The polymorphisms of mucA and fpvA genes correlated well with these profiles, but suggested a relationship between strains isolated from two brothers, not inferred by RFLP. Stop codon mutations in mucA were unique to each dominant strain, indicating the adaptive process suffered. The alternate detection of the same mucA and/or fpvA genotypic variants suggested the coexistence of several subpopulations. This hypothesis was confirmed in a prospective study in which 6 variants were isolated in 7 days from the same patient. CONCLUSIONS: Genotypic variants of the P. aeruginosa dominant strains can coexist in the chronic colonization in CF patients. These variants can be undetected by RFLP and they might present variable antibiotic susceptibility.
Assuntos
Proteínas da Membrana Bacteriana Externa/genética , Proteínas de Bactérias/genética , Fibrose Cística/microbiologia , Polimorfismo Genético , Pseudomonas aeruginosa/genética , Adolescente , Adulto , Criança , Feminino , Variação Genética , Humanos , Masculino , Adulto JovemRESUMO
Introducción Pseudomonas aeruginosa coloniza la mucosa respiratoria del paciente con fibrosis quística (FQ), seleccionándose en general una única cepa dominante en un proceso de variación genética. Métodos Se obtuvieron secuencialmente 123 aislados de P. aeruginosa en 6 pacientes con FQ atendidos durante su seguimiento habitual o en las exacerbaciones a lo largo de 2-12 años en el Hospital Universitario Ramón y Cajal (Madrid). Otros 13 aislados se obtuvieron de un sólo paciente en un estudio a corto plazo (7 días). Se estudió su sensibilidad antimicrobiana por microdilución y caracterizaron mediante polimorfismo de fragmentos de restricción (RFLP) y secuenciación de los genes mucA y fpvA, que codifican respectivamente el regulador de la síntesis de alginato y un receptor de pioverdina. Resultados Se identificó en cada paciente una cepa dominante según el perfil de RFLP. Los polimorfismos de los genes mucA y fpvA se correlacionaron bien con dicho perfil, pero en dos pacientes hermanos se observó una relación entre cepas no evidente por RFLP. Las mutaciones sin sentido en mucA fueron exclusivas de la cepa dominante de cada paciente, reflejando el proceso adaptativo. La detección alternativa de los mismos polimorfismos en mucA o fpvA mostró la coexistencia de subpoblaciones en cada paciente. Esta hipótesis se confirmó en el estudio prospectivo de 7 días al aislar 6 variantes en un único paciente. Conclusiones En la colonización crónica por P. aeruginosa en pacientes con FQ coexisten variantes genotípicas no siempre detectadas por RFLP y con diferentes perfiles de sensibilidad (AU)
Introduction Pseudomonas aeruginosa is able to colonize the lungs of cystic fibrosis patients (CF) in an adaptive process that results in the selection of a dominant strain through a process of genetic variation. Methods One hundred and twenty tree isolates of P. aeruginosa were sequentially recovered from 6 CF patients during the routine follow-up or exacerbations over periods of 2 to 12 years in the Ramon y Cajal University Hospital (Madrid, Spain). Another 13 isolates were obtained from a single CF patient in a short-term study. They were analysed by restriction fragment length polymorphism (RFLP) and sequencing of mucA and fpvA genes, which code for the alginate biosynthesis regulator and a pyoverdin receptor, respectively, and their antibiotic susceptibility was studied by microdilution. Results A dominant colonising strain was found in each patient based on the RFLP profile. The polymorphisms of mucA and fpvA genes correlated well with these profiles, but suggested a relationship between strains isolated from two brothers, not inferred by RFLP. Stop codon mutations in mucA were unique to each dominant strain, indicating the adaptive process suffered. The alternate detection of the same mucA and/or fpvA genotypic variants suggested the coexistence of several subpopulations. This hypothesis was confirmed in a prospective study in which 6 variants were isolated in 7 days from the same patient. Conclusions Genotypic variants of the P. aeruginosa dominant strains can coexist in the chronic colonization in CF patients. These variants can be undetected by RFLP and they might present variable antibiotic susceptibility (AU)
Assuntos
Humanos , Fibrose Cística/complicações , Pseudomonas aeruginosa/genética , Infecções Respiratórias/microbiologia , Polimorfismo de Fragmento de Restrição/genética , /métodosRESUMO
Pseudomonas aeruginosa is isolated in sputum cultures from cystic fibrosis (CF) patients and adults with bronchiectasis (BS) and chronic obstructive pulmonary disease, but it is not well known if the characteristics of colonization in these latter patients are similar to those with CF. We examined 125 P. aeruginosa isolates obtained from 31 patients suffering from these diseases by pulsed field gel electrophoresis and genotyping of mucA and fpvA genes. The pattern of colonization, with dominance of a clonal strain and incidence of mucoid phenotypes, was similar in every group of patients; however, in some CF and BS patients, we detected the replacement or coexistence of 2 main clones. The main differences were found in the nucleotide position of less common mucA mutations, other than mucA22, and in the predominance of the different types of the pyoverdine receptor. Our results support a similar colonization pattern by P. aeruginosa in the different obstructive pulmonary diseases.
