title: A comparison of methods for extracting DNA from Coxiella burnetii as measured by a duplex qPCR assay.
abstract: OBJECTIVE
  To determine the optimal DNA extraction method for the detection of Coxiella burnetii including the small-cell variant (SCV) by real-time PCR (qPCR) in clinical samples.
  
  
  RESULTS
  A duplex qPCR detecting two Coxiella burnetii gene targets (com1 and IS1111a genes) was developed. Each target in this PCR had a sensitivity of one copy number per reaction. DNA extraction methods were compared on spiked negative samples and included a silica column kit, a chloroform separation prior to a silica column method and a chloroform/phenol separation and DNA precipitation method.
  
  
  CONCLUSIONS
  The silica column extraction method was more efficient at recovering C. burnetii DNA, from large-cell and small-cell variants, than a chloroform or chloroform/phenol method. The silica column method was useful on spiked human samples including serum, buffy coat and bone marrow samples.
  
  
  CONCLUSIONS
  This study demonstrated that a simple column kit method is efficient to use for the detection of C. burnetii in clinical samples including the SCV.
authors.affiliation: The Australian Rickettsial Reference Laboratory/Barwon Biomedical Research, The Geelong Hospital, Bellarine Street, Geelong, Victoria 3220, Australia.
authors.forename: M G
authors.initials: MG
authors.lastname: Lockhart
authors.forename: S R
authors.initials: SR
authors.lastname: Graves
authors.forename: M J
authors.initials: MJ
authors.lastname: Banazis
authors.forename: S G
authors.initials: SG
authors.lastname: Fenwick
authors.forename: J
authors.initials: J
authors.lastname: Stenos
mesh_terms: D016997:Coxiella burnetii / Q000235:genetics*; D004269:DNA, Bacterial / Q000737:chemistry* / Q000235:genetics; D006801:Humans; D008967:Molecular Biology / Q000379:methods*; D019800:Phenol / Q000737:chemistry; D016133:Polymerase Chain Reaction; D012822:Silicon Dioxide / Q000737:chemistry
chemical_list: D004269:DNA, Bacterial; D019800:Phenol; D012822:Silicon Dioxide