Abstract
Coxiella burnetii is the causative agent of Q fever, a zoonosis infecting domestic ruminants and humans. Currently used routine diagnostic tools offer limited sensitivity and specificity and symptomless infected animals may be missed. Therefore, diagnostic tools of higher sensitivity and specificity must be developed. For this purpose, the C. burnetii outer membrane protein Com1 was cloned and expressed in Escherichia coli. The His-tagged recombinant protein was purified and used in an indirect enzyme-linked immunosorbent assay (ELISA). Assay performance was tested with more than 400 positive and negative sera from sheep, goats and cattle from 36 locations. Calculation of sensitivity and specificity was undertaken using receiver operating characteristic (ROC) curves. The sensitivities and specificities for sheep were 85% and 68% (optical density at 450nm, OD450 cut-off value 0.32), for goats 94% and 77% (OD450 cut-off value 0.23) and for cattle 71% and 70% (OD450 cut-off value 0.18), respectively. These results correspond to excellent, outstanding and acceptable discrimination of positive and negative sera. In summary, recombinant Com1 can provide a basis for more sensitive and specific diagnostic tools in veterinary medicine.
Highlights
Coxiella burnetii, the etiological agent of Q fever, is an intracellular, Gram-negative bacterium that causes local outbreaks in man and animals worldwide almost every year [1,2,3]
The former serum was from a defined outbreak, where only two samples showed negative results in the commercial enzyme-linked immunosorbent assay (ELISA)
The latter serum was from another defined outbreak and the only one having a negative result in the commercial ELISA from this location
Summary
The etiological agent of Q fever, is an intracellular, Gram-negative bacterium that causes local outbreaks in man and animals worldwide almost every year [1,2,3]. Domestic ruminants are considered as main reservoir hosts and infections in particular in sheep and goats are associated with human cases [4,5,6]. As reproductive disorders can have different etiologies, Q fever is often not considered and the spread of C. burnetii via aerosols proceeds [9]. C. burnetii is highly contagious for humans and is an occupational hazard for people in close contact with infected ruminants like farmers, shepherds and veterinarians as well as abattoir workers [5,6,10]. More than 4000 acute and 284 chronic human cases were reported, mirrored by high numbers of seropositive blood donors [15,16]
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