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Journal

2008 | 3 | 2 | 157-162

Article title

Comparason of extraction methods for PCR detection of Burkholderia cepacia complex (BCC) from cystic fibrosis patients

Content

Title variants

Languages of publication

EN

Abstracts

EN
Direct detection of Burkholderia cepacia complex (BCC) and its genomovars from sputum by molecular tests emerges as a method for rapid identification. In this study, four DNA extraction methods were evaluated for the identification for BCC from sputum of CF patients. Sputa from 28 CF patients were aliquoted and spiked with BCC reference strain. Boiling, phenol-chloroform, CTAB methods and a commercial spin column kit was used for DNA extraction. Total DNA yields were determined by spectrophotometry and single-round recA PCR was used for detection of BCC. No significant difference was observed in DNA yields from different extraction methods. Lower limit of detection for recA PCR was determined as 106 cfu/ml. Amplification was observed in 7/16 (43.7%) of sputa for boiling, 8/16 (50%) of sputa for CTAB and 13/16 (81.2%) of sputa for phenol-chloroform method and spin column kit in the assay sensitivity range determined in the study. Phenol-chloroform and commercial spin column kit were found to be better suited for DNA purification from sputum of CF patients for BCC identification. Diagnostic impact of single-round recA PCR directly from sputum was limited to chronically-infected patients.

Publisher

Journal

Year

Volume

3

Issue

2

Pages

157-162

Physical description

Dates

published
1 - 6 - 2008
online
9 - 4 - 2008

Contributors

author
  • Department of Microbiology and Clinical Microbiology, Hacettepe University Faculty of Medicine, Morphology Building 3rd floor, Sihhiye, Ankara, 06100, Turkey
  • Department of Microbiology and Clinical Microbiology, Hacettepe University Faculty of Medicine, Morphology Building 3rd floor, Sihhiye, Ankara, 06100, Turkey
author
  • Department of Microbiology and Clinical Microbiology, Hacettepe University Faculty of Medicine, Morphology Building 3rd floor, Sihhiye, Ankara, 06100, Turkey
author
  • Department of Pediatrics, Division of Chest Diseases; Ihsan Dogramaci Childrens’ Hospital, Hacettepe University Faculty of Medicine, Sihhiye, Ankara, 06100, Turkey
  • Department of Biostatistics, Hacettepe University Faculty of Medicine, Sihhiye, Ankara, 06100, Turkey
author
  • Department of Pediatrics, Division of Chest Diseases; Ihsan Dogramaci Childrens’ Hospital, Hacettepe University Faculty of Medicine, Sihhiye, Ankara, 06100, Turkey

References

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Document Type

Publication order reference

Identifiers

YADDA identifier

bwmeta1.element.-psjd-doi-10_2478_s11536-007-0069-4
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