This Article
Right arrow Full Text
Right arrow Full Text (PDF)
Right arrow Supplemental material
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrowReprints and Permissions
Right arrow Copyright Information
Right arrow Books from ASM Press
Right arrow MicrobeWorld
Google Scholar
Right arrow Articles by Coscollá, M.
Right arrow Articles by González-Candelas, F.
PubMed
Right arrow PubMed Citation
Right arrow Articles by Coscollá, M.
Right arrow Articles by González-Candelas, F.

 Previous Article  |  Next Article 

Journal of Clinical Microbiology, September 2009, p. 2901-2905, Vol. 47, No. 9
0095-1137/09/$08.00+0     doi:10.1128/JCM.00268-09
Copyright © 2009, American Society for Microbiology. All Rights Reserved.

Direct Sequencing of Legionella pneumophila from Respiratory Samples for Sequence-Based Typing Analysis{triangledown} ,{dagger}

Mireia Coscollá* and Fernando González-Candelas

Institut Cavanilles de Biodiversitat i Biologia Evolutiva and Departament de Genètica, Universitat de València, CIBER Epidemiología y Salud Pública (CIBERESP), Área Genómica y Salud, Centro Superior de Investigación en Salud Pública, Valencia, Spain

Received 6 February 2009/ Returned for modification 1 April 2009/ Accepted 10 July 2009

We have developed a procedure to test the efficiency and reliability of sequencing of Legionella pneumophila genes directly from respiratory samples and have compared the results with those derived from cultured isolates. We tried to obtain the nucleotide sequences of six protein-coding loci included in the sequence-based typing scheme for Legionella pneumophila and three intergenic regions from 132 samples corresponding to 106 patients positive for urine antigen. A seminested PCR approach was used to amplify and sequence these nine loci directly from respiratory samples. Nucleotide sequences were directly obtained for 23 Legionella isolates and also for 66 respiratory secretions from a total of 69 patients. The efficiency of sequencing from respiratory secretions was higher than that of sequencing after the isolation of the Legionella isolates. Moreover, the perfect match between the sequences obtained by both approaches when respiratory samples and cultured isolates from the same patient were available corroborates the suitability of the direct sequencing approach for the identification of Legionella species and molecular epidemiology studies with Legionella species.

* Corresponding author. Mailing address: Institut Cavanilles de Biodiversitat i Biologia Evolutiva, University of Valencia, Apartado Oficial 22085, Valencia 46071, Spain. Phone: 34 963 543 648. Fax: 34 963 543 670. E-mail: mireia.coscolla{at}uv.es

{triangledown} Published ahead of print on 15 July 2009.

{dagger} Supplemental material for this article may be found at http://jcm.asm.org/.

Journal of Clinical Microbiology, September 2009, p. 2901-2905, Vol. 47, No. 9
0095-1137/09/$08.00+0     doi:10.1128/JCM.00268-09
Copyright © 2009, American Society for Microbiology. All Rights Reserved.





Copyright © 2009 by the American Society for Microbiology.   For an alternate route to Journals.ASM.org, visit: http://intl-journals.asm.org | More Info»