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Journal of Clinical Microbiology, October 2005, p. 5136-5142, Vol. 43, No. 10
0095-1137/05/$08.00+0     doi:10.1128/JCM.43.10.5136-5142.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.

Transmission of Burkholderia cepacia Complex: Evidence for New Epidemic Clones Infecting Cystic Fibrosis Patients in Italy

S. Campana,^1* G. Taccetti,¹ N. Ravenni,¹ F. Favari,⁴ L. Cariani,⁴ A. Sciacca,⁶ D. Savoia,⁷ A. Collura,⁸ E. Fiscarelli,⁹ G. De Intinis,¹⁰ M. Busetti,¹¹ A. Cipolloni,¹² A. d'Aprile,¹³ E. Provenzano,¹⁴ I. Collebrusco,¹⁵ P. Frontini,¹⁶ G. Stassi,¹⁷ M. Trancassini,¹⁸ D. Tovagliari,¹⁹ A. Lavitola,²⁰ C. J. Doherty,³ T. Coenye,² J. R. W. Govan,³ and P. Vandamme²

Department of Paediatrics, Cystic Fibrosis Center, University of Florence, Florence, Italy,¹ Laboratorium voor Microbiologie, Ghent University, Ghent, Belgium,² Medical Microbiology, Centre for Infectious Diseases, University of Edinburgh, Edinburgh, United Kingdom,³ Ospedale Civile Maggiore, Verona, Italy,⁴ Istituti Clinici di Perfezionamento, Milan, Italy,⁵ Università di Catania, Catania, Italy,⁶ Università di Torino, Turin, Italy,⁷ Ospedale dei Bambini G. de Cristina, Palermo, Italy,⁸ Ospedale Bambin Gesù, Rome, Italy,⁹ Azienda Ospedaliera OIRM S. Anna, Turin, Italy,¹⁰ Istituto di Igiene, Trieste, Italy,¹¹ Ospedale M. Bufalini, Cesena, Italy,¹² Ospedale G. Tatarella, Cerignola, Italy,¹³ Divisione Pediatrica, Ospedale di Soverato, Soverato, Italy,¹⁴ Ospedale Calai, Gualdo Tadino, Italy,¹⁵ Ospedale G. Salesi, Ancona, Italy,¹⁶ Azienda Ospedaliera Universitaria di Messina, Messina, Italy,¹⁷ Università la Sapienza, Rome, Italy,¹⁸ Università di Parma, Parma, Italy,¹⁹ Università di Napoli, Naples, Italy,²⁰

Received 3 March 2005/ Returned for modification 30 April 2005/ Accepted 17 July 2005

To analyze national prevalence, genomovar distribution, and epidemiology of the Burkholderia cepacia complex in Italy, 225 putative B. cepacia complex isolates were obtained from 225 cystic fibrosis (CF) patients attending 18 CF centers. The genomovar status of these isolates was determined by a polyphasic approach, which included whole-cell protein electrophoresis and recA restriction fragment length polymorphism (RFLP) analysis. Two approaches were used to genotype B. cepacia complex isolates: BOX-PCR fingerprinting and pulsed-field gel electrophoresis (PFGE) of genomic macrorestriction fragments. A total of 208 (92%) of 225 isolates belonged to the B. cepacia complex, with Burkholderia cenocepacia as the most prevalent species (61.1%). Clones delineated by PFGE were predominantly linked to a single center; in contrast, BOX-PCR clones were composed of isolates collected either from the same center or from different CF centers and comprised multiple PFGE clusters. Three BOX-PCR clones appeared of special interest. One clone was composed of 17 B. cenocepacia isolates belonging to recA RFLP type H. These isolates were collected from six centers and represented three PFGE clusters. The presence of insertion sequence IS1363 in all isolates and the comparison with PHDC reference isolates identified this clone as PHDC, an epidemic clone prominent in North American CF patients. The second clone included 22 isolates from eight centers and belonged to recA RFLP type AT. The genomovar status of strains with the latter RFLP type is not known. Most of these isolates belonged to four different PFGE clusters. Finally, a third clone comprised nine B. pyrrocinia isolates belonging to recA RFLP type Se13. They represented three PFGE clusters and were collected in three CF centers.

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* Corresponding author. Mailing address: Cystic Fibrosis Center, Department of Pediatrics, Anna Meyer Children's Hospital, University of Florence, 50132 Florence, Italy. Phone: 39 055 5662509. Fax: 39 055 5662474. E-mail: s.campana{at}meyer.it.

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Journal of Clinical Microbiology, October 2005, p. 5136-5142, Vol. 43, No. 10
0095-1137/05/$08.00+0     doi:10.1128/JCM.43.10.5136-5142.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.

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