Murdoch University Research Repository

Welcome to the Murdoch University Research Repository

The Murdoch University Research Repository is an open access digital collection of research
created by Murdoch University staff, researchers and postgraduate students.

Learn more

A comparison of methods for extracting DNA from Coxiella burnetii as measured by a duplex qPCR assay

Lockhart, M.G., Graves, S.R., Banazis, M.J., Fenwick, S.G. and Stenos, J. (2011) A comparison of methods for extracting DNA from Coxiella burnetii as measured by a duplex qPCR assay. Letters in Applied Microbiology, 52 (5). pp. 514-520.

Free to read:
*No subscription required


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.
Methods and 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.
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.
Significance and impact of study:
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.

Item Type: Journal Article
Murdoch Affiliation(s): School of Veterinary and Biomedical Sciences
Publisher: Blackwell Publishing
Copyright: 2011 The Authors
Item Control Page Item Control Page