Journal article
A comparison of methods for extracting DNA from Coxiella burnetii as measured by a duplex qPCR assay
Letters in Applied Microbiology, Vol.52(5), pp.514-520
2011
Abstract
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.
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.
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.
Details
- Title
- A comparison of methods for extracting DNA from Coxiella burnetii as measured by a duplex qPCR assay
- Authors/Creators
- M.G. Lockhart (Author/Creator) - Murdoch UniversityS.R. Graves (Author/Creator)M.J. Banazis (Author/Creator) - Murdoch UniversityS.G. Fenwick (Author/Creator)J. Stenos (Author/Creator) - Geelong Hospital
- Publication Details
- Letters in Applied Microbiology, Vol.52(5), pp.514-520
- Publisher
- Blackwell Publishing
- Identifiers
- 991005546035607891
- Copyright
- 2011 The Authors
- Murdoch Affiliation
- School of Veterinary and Biomedical Sciences
- Language
- English
- Resource Type
- Journal article
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- Collaboration types
- Domestic collaboration
- Citation topics
- 1 Clinical & Life Sciences
- 1.258 Zoonotic Diseases
- 1.258.2263 Coxiella Burnetii
- Web Of Science research areas
- Biotechnology & Applied Microbiology
- Microbiology
- ESI research areas
- Biology & Biochemistry