title: Polyphasic characterisation of Burkholderia cepaciacomplex species isolated from children with cystic fibrosis.
abstract: Cystic fibrosis (CF) patients with Burkholderia cepacia complex (Bcc) pulmonary infections have high morbidity and mortality. The aim of this study was to compare different methods for identification of Bcc species isolated from paediatric CF patients. Oropharyngeal swabs from children with CF were used to obtain isolates of Bcc samples to evaluate six different tests for strain identification. Conventional (CPT) and automatised (APT) phenotypic tests, polymerase chain reaction (PCR)-recA, restriction fragment length polymorphism-recA, recA sequencing, and matrix-assisted laser desorption ionization-time of flight (MALDI-TOF) were applied. Bacterial isolates were also tested for antimicrobial susceptibility. PCR-recA analysis showed that 36 out of the 54 isolates were Bcc. Kappa index data indicated almost perfect agreement between CPT and APT, CPT and PCR-recA, and APT and PCR-recA to identify Bcc, and MALDI-TOF and recA sequencing to identify Bcc species. The recA sequencing data and the MALDI-TOF data agreed in 97.2% of the isolates. Based on recA sequencing, the most common species identified were Burkholderia cenocepacia IIIA (33.4%),Burkholderia vietnamiensis (30.6%), B. cenocepaciaIIIB (27.8%), Burkholderia multivorans (5.5%), and B. cepacia (2.7%). MALDI-TOF proved to be a useful tool for identification of Bcc species obtained from CF patients, although it was not able to identify B. cenocepacia subtypes.
authors.affiliation: Laboratório Municipal de Curitiba, Curitiba, PR, Brasil.
authors.forename: Fernando José
authors.initials: FJ
authors.lastname: Vicenzi
authors.affiliation: Departamento de Microbiologia, Escola de Saúde e Biociências, Pontifícia Universidade Católica do Paraná, Curitiba, PR, Brasil.
authors.forename: Marcelo
authors.initials: M
authors.lastname: Pillonetto
authors.affiliation: Hospital de Clínicas, Faculdade de Medicina, Universidade Federal do Paraná, Curitiba, PR, Brasil.
authors.forename: Helena Aguilar Peres Homem de Mello de
authors.initials: HA
authors.lastname: Souza
authors.affiliation: Instituto de Pesquisa Pelé Pequeno Principe, Faculdades Pequeno Príncipe, Curitiba, PR, Brasil.
authors.forename: Jussara Kasuko
authors.initials: JK
authors.lastname: Palmeiro
authors.affiliation: Departamento de Pediatria, Faculdade de Medicina, Universidade Federal do Paraná, Curitiba, PR, Brasil.
authors.forename: Carlos Antônio
authors.initials: CA
authors.lastname: Riedi
authors.affiliation: Departamento de Pediatria, Faculdade de Medicina, Universidade Federal do Paraná, Curitiba, PR, Brasil.
authors.forename: Nelson Augusto
authors.initials: NA
authors.lastname: Rosario-Filho
authors.affiliation: Instituto de Pesquisa Pelé Pequeno Principe, Faculdades Pequeno Príncipe, Curitiba, PR, Brasil.
authors.forename: Libera Maria
authors.initials: LM
authors.lastname: Dalla-Costa
mesh_terms: D001426:Bacterial Proteins / Q000235:genetics; D015373:Bacterial Typing Techniques; D019121:Burkholderia Infections / Q000821:virology*; D042602:Burkholderia cepacia complex / Q000145:classification / Q000235:genetics*; D002648:Child; D002675:Child, Preschool; D003550:Cystic Fibrosis / Q000821:virology*; D004269:DNA, Bacterial / Q000235:genetics; D005260:Female; D006801:Humans; D007223:Infant; D008297:Male; D009960:Oropharynx / Q000821:virology; D010641:Phenotype; D016133:Polymerase Chain Reaction; D012150:Polymorphism, Restriction Fragment Length; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
chemical_list: D001426:Bacterial Proteins; D004269:DNA, Bacterial
references.citation: J Clin Microbiol. 2012 Dec;50(12):3845-52
references.pmid: 22993178
references.citation: Syst Appl Microbiol. 2011 Apr;34(2):87-95
references.pmid: 21257278
references.citation: Clin Microbiol Infect. 2010 Jul;16(7):821-30
references.pmid: 20880411
references.citation: Clin Microbiol Infect. 2010 Jul;16(7):809-20
references.pmid: 20880410
references.citation: Clin Microbiol Rev. 2010 Apr;23(2):299-323
references.pmid: 20375354
references.citation: Int J Syst Evol Microbiol. 2009 Jan;59(Pt 1):102-11
references.pmid: 19126732
references.citation: Eur J Clin Microbiol Infect Dis. 2008 Nov;27(11):1101-11
references.pmid: 18600352
references.citation: FEMS Microbiol Lett. 2005 May 1;246(1):39-45
references.pmid: 15869960
references.citation: J Pediatr (Rio J). 2006 Sep-Oct;82(5):377-82
references.pmid: 17003938
references.citation: Int J Syst Evol Microbiol. 2008 Jul;58(Pt 7):1580-90
references.pmid: 18599699
references.citation: J Clin Microbiol. 1997 Mar;35(3):614-9
references.pmid: 9041399
references.citation: Biometrics. 1977 Mar;33(1):159-74
references.pmid: 843571
references.citation: Eur Respir J. 2004 Jun;23(6):851-6
references.pmid: 15218997
references.citation: J Appl Microbiol. 2003;95(6):1191-9
references.pmid: 14632991
references.citation: J Med Microbiol. 2002 Nov;51(11):937-40
references.pmid: 12448677
references.citation: FEMS Microbiol Lett. 2002 Aug 27;214(1):1-5
references.pmid: 12204364
references.citation: J Med Microbiol. 2002 Jul;51(7):533-8
references.pmid: 12132768
references.citation: J Clin Microbiol. 2001 Oct;39(10):3427-36
references.pmid: 11574551
references.citation: Am J Respir Crit Care Med. 2001 Jul 1;164(1):92-6
references.pmid: 11435245
references.citation: J Clin Microbiol. 2000 Sep;38(9):3165-73
references.pmid: 10970351
references.citation: J Appl Microbiol. 2008 Jun;104(6):1539-51
references.pmid: 18217926
references.citation: Int J Syst Evol Microbiol. 2015 Jul;65(7):2265-71
references.pmid: 25872960
references.citation: Pol J Microbiol. 2013;62(4):373-6
references.pmid: 24730131
references.citation: Diagn Microbiol Infect Dis. 2013 Oct;77(2):126-8
references.pmid: 23891221
references.citation: Syst Appl Microbiol. 2013 Oct;36(7):483-9
references.pmid: 23867250