[Use of 16S rRNA gene sequencing for identification of "Pseudomonas-like" isolates from sputum of patients with cystic fibrosis]. / Mucoviscidose: identification des bacilles isolés d'infections bronchiques par séquençage du gène de l'ARN ribosomal 16S.
Pathol Biol (Paris)
; 53(8-9): 500-2, 2005.
Article
en Fr
| MEDLINE
| ID: mdl-16081224
Since nonfermenting, Gram negative bacilli recovered from patients with cystic fibrosis could be misidentified with phenotypic procedures, we used partial 16S ribosomal RNA gene (16S gene) sequencing to identify these "Pseudomonas-like" isolates. 473 isolates were recovered from 66 patients in 2003. Sequencing was used to identify 29 (from 24 patients) of the 473 isolates, showing unclear results with routine tests. PCR with specific primers was carried out to amplify a 995 bp fragment, which was then sequenced. The sequences were analyzed with GenBank database for species assignment. Phenotypic and genotypic results were concordant for 20/29 isolates (10 Pseudomonas aeruginosa, 5 Burkholderia cepacia, 3 Stenotrophomonas maltophilia, 2 Achromobacter xylosoxidans). However, 3 of the 5 B. cepacia isolates were then identified as Burkholderia multivorans with a PCR-RFLP procedure. Phenotypic misidentification was observed for 9/29 isolates: 4 A. xylosoxidans, 1 P. aeruginosa, 1 Bordetella petrii, 1 Bordetella bronchiseptica, 1 Ralstonia respiraculi and 1 Ralstonia mannitolilytica. Partial 16S gene sequencing improved the identification of "Pseudomonas-like" isolates from cystic fibrosis patients, but the accuracy to distinguish between genomovars of the B. cepacia complex was inadequate.
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Colección:
01-internacional
Base de datos:
MEDLINE
Asunto principal:
Pseudomonas
/
Esputo
/
ARN Bacteriano
/
ARN Ribosómico 16S
/
Fibrosis Quística
Tipo de estudio:
Diagnostic_studies
Límite:
Humans
Idioma:
Fr
Revista:
Pathol Biol (Paris)
Año:
2005
Tipo del documento:
Article
País de afiliación:
Francia
Pais de publicación:
Francia