Molecular Identification of Burkholderia Cepacia Complex and Species Distribution Among Cystic Fibrosis Patients Seen at the Reference Center in Southern Brazil

Autores

  • Fernanda Concli Leite Hospital de Clínicas de Porto Alegre, RS
  • Alice Beatriz Mombach Pinheiro Machado Hospital de Clínicas de Porto Alegre
  • Larissa Lutz Hospital de Clínicas de Porto Alegre
  • Maria Izolete Vieira Hospital de Clínicas de Porto Alegre
  • Afonso Luis Barth Hospital de Clínicas de Porto Alegre, Universidade Federal do Rio Grande do Sul

Palavras-chave:

Burkholderia cepacia, fibrose cística, PCR-RFLP, gene recA

Resumo

Background: Burkholderia cepacia complex (Bcc) infections in cystic fibrosis (CF) patients are associated with decline in lung function and reduced survival. The potential transmissibility of Bcc among CF patients has been reported, indicating that strict segregation of CF patients with Bcc is crucial.

Aims: To standardize the PCR-RFLP (polymerase chain reaction-restriction fragment length polymorphism) assay in order to identify Bcc species and to establish the prevalence of Bcc species and their susceptibility profile among CF patients seen at the Hospital de Clínicas de Porto Alegre (HCPA).

Methods: The classification of the clinical isolates recovered from respiratory tract specimens of CF patients as Bcc was achieved using the API-20NE® phenotypic commercial system. The identification of the Bcc species was performed using PCR-RFLP. The antimicrobial disk diffusion susceptibility testing was performed according to the CLSI (2006).

Results: API-20NE® was able to identify Bcc isolates (244 specimens), such as B. cepacia, indicating that it was not able to distinguish among the Bcc species. The PCR-RFLP molecular method discriminated the eight reference Bcc species, thus validating the method for clinical isolates. Bcc prevalence determined by PCR-RFLP was 10.6% (26/244). The molecular analysis identified B. cenocepacia in 53.8% (14/26) of infected patients, B. multivorans in 15.4% (4/26), and B. vietnamiensis and B. ambifaria in 7.7% (2/26). The antibiotic resistance profile was variable among Bcc species.

Conclusions: The PCR-RFLP method was validated for the identification of Bcc species. B. cenocepacia proved to be the most prevalent species among the CF patients seen at the HCPA.

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Biografia do Autor

Alice Beatriz Mombach Pinheiro Machado, Hospital de Clínicas de Porto Alegre

Unidade de Biologia Molecular, Serrviço de Patologia  Clínica

Larissa Lutz, Hospital de Clínicas de Porto Alegre

Unidade de Microbiologia, Serviço de Patologia Clínica

Maria Izolete Vieira, Hospital de Clínicas de Porto Alegre

Unidade de Microbiologia, Serviço de Patologia Clínica

Afonso Luis Barth, Hospital de Clínicas de Porto Alegre, Universidade Federal do Rio Grande do Sul

Programa de Pós -graduação em Medicina, Serviço de Patologia Clínica

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Publicado

2011-07-27

Como Citar

1.
Leite FC, Pinheiro Machado ABM, Lutz L, Vieira MI, Barth AL. Molecular Identification of Burkholderia Cepacia Complex and Species Distribution Among Cystic Fibrosis Patients Seen at the Reference Center in Southern Brazil. Clin Biomed Res [Internet]. 27º de julho de 2011 [citado 4º de dezembro de 2022];31(2). Disponível em: https://seer.ufrgs.br/index.php/hcpa/article/view/21153

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