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Journal of Clinical Microbiology, June 1998, p. 1642-1645, Vol. 36, No. 6
0095-1137/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.
Comparison of Culture and PCR for Detection of Burkholderia
cepacia in Sputum Samples of Patients with Cystic
Fibrosis
Paul W.
Whitby,1
Hani L. N.
Dick,2
Preston W.
Campbell III,3
D. Elizabeth
Tullis,2
Anne
Matlow,4 and
Terrence
L.
Stull1,5,*
Departments of
Pediatrics1 and
Microbiology/Immunology,5 University of
Oklahoma Health Sciences Center, Oklahoma City, Oklahoma 73104;
The Wellesley Hospital, Toronto, Ontario M4Y
1J3,2 and
The Hospital for Sick
Children, Toronto, Ontario M5G 1X8,4 Canada; and
Division of Pediatric Pulmonology, Vanderbilt University
School of Medicine, Nashville, Tennessee 372323
Received 16 October 1997/Returned for modification 2 February
1998/Accepted 13 March 1998
We investigated the utility of PCR to detect Burkholderia
cepacia directly in sputum samples at two cystic fibrosis (CF)
centers serving children and adults. Following liquefaction of the
sputa by using N-acetyl-L-cysteine, DNA was
isolated and analyzed by PCRs with three different primer pairs
directed toward bacterial rRNA loci. Two primer pairs were putatively
specific for B. cepacia. The other pair, which
universally amplifies a band from all bacteria, served as a control.
Sputum samples were obtained from 219 patients and analyzed
independently by culture and by PCR to detect B. cepacia. The analyses were performed blinded with respect to each other. The results of the PCR with sputa demonstrated that the primers
directed to the 16S loci demonstrated approximately 95% concordance
with culture results and were more specific than those amplifying the
16S to 23S spacer region. In addition, the 16S primer pair putatively
identified B. cepacia in seven patients whose sputa
were culture negative at this time. Of these culture-negative patients,
five had sputum samples that were culture positive for B. cepacia either prior or subsequent to this study. The results of
this study indicate the utility of PCR as a diagnostic method for the
rapid identification of B. cepacia in sputum samples
of CF patients. We anticipate that improvements in our taxonomic understanding may allow the design of more specific primers for detection of each species of the B. cepacia complex in
sputum samples.
*
Corresponding author. Mailing address: Department of
Pediatrics, CHO 2B300, 940 N.E. 13th St., Oklahoma City, OK 73104. Phone: (405) 271-4401. Fax: (405) 271-8710. E-mail:
Terry-Stull{at}uokhsc.edu.
Journal of Clinical Microbiology, June 1998, p. 1642-1645, Vol. 36, No. 6
0095-1137/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.
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