Detection of Burkholderia cepacia DNA from artificially infected EDTA-blood and lung tissue comparing different DNA isolation methods.
Bacterial DNA (Burkholderia cepacia) was prepared from artificially infected equine ethylenediaminetetraacetic acid (EDTA)-blood and lung tissue by using four standard methods (lysis buffer containing proteinase K, phenol/chloroform/isoamylalcohol-extraction, microwave-treatment, heat treatment) and six commercially available kits (Puregene, High Pure PCR Template Preparation Kit, InstaGene, QiaAmp Tissue Kit, DNAzol and Elu-Quik). After a subsequent polymerase chain reaction (PCR), their efficacy and sensitivity were compared. Concerning the detection limits, the simple lysis with a proteinase K-containing buffer led to the best results for EDTA-blood as well as for artificially infected lung tissue.
Merk
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Institut fuer Medizinische Informatik und Biomathematik, Domagkstrasse 9, 48149 Munster, Germany.
Meyer
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Greiser-Wilke
I
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Sprague
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LD
Neubauer
H
H
D000818:Animals; D019121:Burkholderia Infections / Q000175:diagnosis / Q000382:microbiology / Q000662:veterinary; D016956:Burkholderia cepacia / Q000302:isolation & purification*; D004269:DNA, Bacterial / Q000032:analysis*; D004492:Edetic Acid; D006734:Horse Diseases / Q000175:diagnosis / Q000382:microbiology; D006736:Horses; D008168:Lung / Q000382:microbiology; D016133:Polymerase Chain Reaction / Q000379:methods / Q000662:veterinary*; D012680:Sensitivity and Specificity
D004269:DNA, Bacterial; D004492:Edetic Acid