Detection of Burkholderia cepacia DNA from artificially infected EDTA-blood and lung tissue comparing different DNA isolation methods

J Vet Med B Infect Dis Vet Public Health. 2006 Aug;53(6):281-5. doi: 10.1111/j.1439-0450.2006.00956.x.

Abstract

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.

Publication types

  • Comparative Study

MeSH terms

  • Animals
  • Burkholderia Infections / diagnosis
  • Burkholderia Infections / microbiology
  • Burkholderia Infections / veterinary
  • Burkholderia cepacia / isolation & purification*
  • DNA, Bacterial / analysis*
  • Edetic Acid
  • Horse Diseases / diagnosis
  • Horse Diseases / microbiology
  • Horses
  • Lung / microbiology
  • Polymerase Chain Reaction / methods
  • Polymerase Chain Reaction / veterinary*
  • Sensitivity and Specificity

Substances

  • DNA, Bacterial
  • Edetic Acid