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Detection of Pseudomonas ( Burkholderia ) cepacia using PCR
Author(s) -
Campbell Preston W.,
Phillips John A.,
Heidecker Gwendolyn J.,
Krishnamani M. R. S.,
Zahorchak Robert,
Stull Terrence L.
Publication year - 1995
Publication title -
pediatric pulmonology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.866
H-Index - 106
eISSN - 1099-0496
pISSN - 8755-6863
DOI - 10.1002/ppul.1950200109
Subject(s) - burkholderia , sputum , microbiology and biotechnology , pseudomonas aeruginosa , cystic fibrosis , pseudomonas , burkholderia cepacia complex , medicine , polymerase chain reaction , pseudomonadaceae , bacteria , biology , pathology , gene , tuberculosis , genetics , biochemistry
Pseudomonas cepacia colonization of the lung is associated with increased morbidity and mortality for cystic fibrosis (CF) patients. The lack of a sensitive detection method for Pseudomonas cepacia in CF sputum has resulted in controversy regarding its epidemiology. We designed a PCR method to detect P. cepacia using P. cepacia 16 S rRNA sequences as the amplification target region. The PCR amplification with purified DNA as template yielded the expected 209‐bp products from P. cepacia , but not from related Pseudomonas species of medical importance or other bacteria which have been reported to colonize CF patients. In serial dilution experiments as few as 10 2 P. cepacia CFU were detectable. When sputum samples from three CF patients chronically colonized with P. cepacia and P. aeruginosa were analyzed, P. cepacia was detected in all three specimens by PCR, but only in two when selective culture was performed. Our data support the potential role of PCR technology in the rapid, sensitive, and definitive detection of P. cepacia in CF sputum samples, even in the context of concomitant P. aeruginosa colonization. Pediatr Pulmonol. 1995; 20:44–49 . © 1995 Wiley‐Liss, Inc.