title: Real-time PCR for the early detection and quantification of Coxiella burnetii as an alternative to the murine bioassay.
abstract: Real-time PCR was used to analyze archived blood from non-human primates (NHP) 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 bioassay for both quantification and early detection of C. burnetii bacteria. Real-time PCR and the mouse bioassay exhibited no statistical difference in quantifying the number of microorganisms delivered in the aerosol challenge dose. 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. 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.
authors.affiliation: Diagnostic Systems Division, United States Army Medical Research Institute for Infectious Diseases, 1425 Porter St, Frederick, MD 21702, USA.
authors.forename: Gerald B
authors.initials: GB
authors.lastname: Howe
authors.forename: Bonnie M
authors.initials: BM
authors.lastname: Loveless
authors.forename: David
authors.initials: D
authors.lastname: Norwood
authors.forename: Philip
authors.initials: P
authors.lastname: Craw
authors.forename: David
authors.initials: D
authors.lastname: Waag
authors.forename: Marilyn
authors.initials: M
authors.lastname: England
authors.forename: John R
authors.initials: JR
authors.lastname: Lowe
authors.forename: Bernard C
authors.initials: BC
authors.lastname: Courtney
authors.forename: M Louise
authors.initials: ML
authors.lastname: Pitt
authors.forename: David A
authors.initials: DA
authors.lastname: Kulesh
mesh_terms: D000818:Animals; D001681:Biological Assay; D016997:Coxiella burnetii / Q000235:genetics*; D005260:Female; D008252:Macaca fascicularis; D051379:Mice; D016133:Polymerase Chain Reaction / Q000379:methods*; D011778:Q Fever / Q000175:diagnosis*; D015203:Reproducibility of Results