Development of Tools for Surveillance of Coxiella burnetii in Domestic Ruminants and Australian Marsupials and Their Waste
Abstract
The aim of this study was to develop improved methods to detect viable Coxiella burnetii in wastes from livestock production. A quantitative real-time PCR system (qPCR) with high sensitivity and specificity was developed to detect the C. burnetii in environmental samples associated with domestic ruminants and native Australian marsupials. Different detection chemistries and procedures were evaluated based on their sensitivity, specificity and reproducibility. When combined, the IS1111a TaqMan qPCR and Geneworks PowerSoil DNA Extraction Kit provided a test which was capable of detecting as few as two C. burnetii genome equivalents in 0.2g of soil or feces. Coxiella burnetii has been shown to display extreme resistance to environmental exposure. Therefore assessment of the viability of the organism in environmental matrices is more useful for risk assessment programs than detection of DNA alone. A quantitative reverse transcriptase PCR was developed that was able to detect viable C. burnetii cells in soil. The sensitivity of the assay was enhanced by heat-treating the soil samples prior to extraction of RNA. A system was developed to determine the efficacy of various disinfectant treatments against the environmental pathogen C. burnetii. Commercially available ELISA and CFT assays exist for ruminants but there are no immunological tests available for detecting C. burnetii in marsupials even though Australian marsupials are known to be susceptible to C. burnetii. An indirect ELISA for detecting anti-Coxiella antibodies in kangaroos was developed.
Document Details
- Document Type
- Technical Report
- Publication Date
- Jun 12, 2009
- Accession Number
- ADA514412
Entities
People
- Michael Banazis
Organizations
- Murdoch University