Real-time PCR for the Early Detection and Quantification of Coxiella burnetii as an Alternative to the Murine Bioassay
ARMY MEDICAL RESEARCH INST OF INFECTIOUS DISEASES FORT DETRICK MD BACTERIOLOGY DIV
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Real-time PCR was used to analyze archived blood and fluid samples originating from a well-controlled Q fever vaccine efficacy trial. The PCR targets were the IS1111 element and the com1 gene of Coxiella burnetii. Data from that previous study were used to evaluate real-time PCR as an alternative to the use of sero-conversion by mouse bio-assay for both early detection and quantification of C. burnetii bacteria. The presence of C. burnetii in peripheral blood of non-human primates was detected by real-time PCR as early after exposure as the mouse bioassay with results available within hours instead of weeks. In a separate experiment, real-time PCR and the mouse bioassay exhibited no statistical difference in the number of microorganisms delivered in the aerosol challenge dose. Research animals are both costly and subject to stringent regulations so the judicious use of archived samples conserved biological as well as financial resources. This study demonstrates that real-time PCR has the ability to replace the mouse bioassay to measure dosage and monitor infection of C. burnetii in a non-human primate model.