title: Development of a high throughput PCR to detect Coxiella burnetii and its application in a diagnostic laboratory over a 7-year period.
abstract: Q fever is a worldwide zoonotic infectious disease due to Coxiella burnetii. The clinical presentation may be acute (pneumonia and/or hepatitis) or chronic (most commonly endocarditis). Diagnosis mainly relies on serology and PCR. We therefore developed a quantitative real-time PCR. We first tested blindly its performance on various clinical samples and then, when thoroughly validated, we applied it during a 7-year period for the diagnosis of both acute and persistent C. burnetii infection. Analytical sensitivity (< 10 copies/PCR) was excellent. When tested blindly on 183 samples, the specificity of the PCR was 100% (142/142) and the sensitivity was 71% (29/41). The sensitivity was 88% (7/8) on valvular samples, 69% (20/29) on blood samples and 50% (2/4) on urine samples. This new quantitative PCR was then successfully applied for the diagnosis of acute Q fever and endovascular infection due to C. burnetii, allowing the diagnosis of Q fever in six patients over a 7-year period. During a local small cluster of cases, the PCR was also applied to blood from 1355 blood donors; all were negative confirming the high specificity of this test. In conclusion, we developed a highly specific method with excellent sensitivity, which may be used on sera for the diagnosis of acute Q fever and on various samples such as sera, valvular samples, aortic specimens, bone and liver, for the diagnosis of persistent C. burnetii infection.
authors.affiliation: Institute of Microbiology, University Hospital of Lausanne Lausanne, Switzerland.
authors.forename: K
authors.initials: K
authors.lastname: Jaton
authors.affiliation: Infectious Diseases and Immunology, Institut Central des Hôpitaux Valaisans Sion, Switzerland.
authors.forename: O
authors.initials: O
authors.lastname: Peter
authors.affiliation: Faculté de Médecine, Université de la Méditerranée Marseille, 13005, France.
authors.forename: D
authors.initials: D
authors.lastname: Raoult
authors.affiliation: Service Régional Vaudois de Transfusion Sanguine Epalinges, Switzerland.
authors.forename: J-D
authors.initials: JD
authors.lastname: Tissot
authors.affiliation: Institute of Microbiology, University Hospital of Lausanne Lausanne, Switzerland.
authors.forename: G
authors.initials: G
authors.lastname: Greub
keywords: Blood donations; Coxiella burnetii; PCR; Q fever; high throughput