Validation of fluorescence polarization assay (FPA) and comparison with other tests used for diagnosis of B. melitensis infection in sheep.
Fluorescence polarization assay (FPA) is a new test for the serological diagnosis of Brucella spp. infection in animals. The FPA is validated for the diagnosis of B. melitensis infection in sheep. For this purpose, 166 sera originated from natural infected sheep (verified by culture) and 851 sera originated from healthy animals (reared in areas where B. melitensis was never been isolated) were tested. The optimum cut-off value that offers the highest diagnostic sensitivity (DSn) and diagnostic specificity (DSp) was determined at 87mP with the use of ROC analysis. The DSn and DSp of FPA using this cut-off value are calculated at 97.6 and 98.9% with a 95% confidence interval (CI) of 93.9-99.3% and 98.0-99.5%, respectively. The DSn and DSp of FPA have been assessed also using as positive reference (n=587), sera that gave positive results at least in two tests used for diagnosis of B. melitensis in sheep as Rose Bengal Test (RBT), modified Rose Bengal Test (m-RBT), complement fixation test (CFT), indirect Elisa (i-Elisa) and competition Elisa (c-Elisa) originated from animals reared in flocks infected by B. melitensis. The optimum cut-off value using the above panel of positive reference sera was the same offering a DSn of 95.9% with a 95% CI, 94.0-97.4%, since the DSp remains the same. The DSn and DSp as well as performance, accuracy and agreement of FPA's result were compared with those of other tests used. The accuracy of FPA is very high, similar with that of i-Elisa. FPA is a promising assay, which offers a DSn and accuracy better that of those of the tests currently approved for the diagnosis of B. melitensis in sheep and goats. Due to its simplicity, the sort time that results can be obtained and its accuracy it can be used and improve the laboratory testing capacity as well as the efficacy of the eradication program based on test-and-slaughter policy.
Minas
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Technological Educational Institution of Larissa, Faculty of Health Professions, Laboratory of Microbiology, Larissa, 41 110, Greece. aminas@teilar.gr
Stournara
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Minas
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Papaioannou
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Krikelis
V
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Tselepidis
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S
D000818:Animals; D017347:Brucella melitensis / Q000276:immunology*; D002006:Brucellosis / Q000175:diagnosis / Q000453:epidemiology / Q000662:veterinary*; D003937:Diagnosis, Differential; D016231:Fluorescence Polarization Immunoassay / Q000592:standards*; D015511:Goat Diseases / Q000175:diagnosis / Q000453:epidemiology; D006041:Goats; D012372:ROC Curve; D012015:Reference Standards; D012016:Reference Values; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D016036:Seroepidemiologic Studies; D012756:Sheep; D012757:Sheep Diseases / Q000175:diagnosis* / Q000453:epidemiology; D013997:Time Factors