Validation of the Fluorescence Polarization Assay (FPA) for the diagnosis of Bovine Brucellosis

Abstract

Fluorescence polarization assay (FPA), a serological assay, was validated as an alternative test for the rapid and cost-effective diagnosis of bovine brucellosis, with the aim of improving the control of brucellosis in South Africa. The FPA is anticipated to distinguish between vaccinated and infected cattle, circumventing the challenge associated with the tests that are currently used. Positive cattle serum samples (n =420) confirmed by Complement Fixation Test were tested in conjunction with serum samples (n = 446) from non-infected cattle initially tested on Rose Bengal Test, CFT and compared with FPA. The optimum cut-off value that offers the highest diagnostic sensitivity (Dsn) and diagnostic specificity (Dsp) was determined as 87 mP with the use of ROC analysis. The Dsn and Dsp of FPA using this cut-off value was calculated at 99.09% - 100% and 68.09%- 76.61% respectively with a 95% confidence interval (cl). The area under curve (AUC) was calculated at 0.9842 with a 95% standard error (S.E) of 0.005532 with positive and negative likelihood ratio (+LR) (-LR) at 3.643 and 1.002, respectively. The FPA was found to be as effective as CFT and should be considered because of its accuracy and other advantages such as speed, high throughput and the objectivity of the interpretation of results that can be obtained electronically by the (PHERAstar) machine. The test should be included in routine serological diagnosis for brucellosis.

I- Fluorescence polarization assay (i-FPA) ukuhlolwa kwe-serological okuqinisekiswe njengenye indlela yokuhlola ukuxilongwa okusheshayo nengabizi kwe-bovine brucellosis, okuzokwenza ngcono ukulawulwa kwe-brucellosis eNingizimu Afrika. Ngaphezu kwalokho, i-FPA kulindeleke ukuthi yehlukanise phakathi kwezinkomo ezigonyiwe nezithelelekile futhi lokhu kuzonciphisa inselelo ehambisana nokuhlolwa esetshenziswa njengamanje. Amasampula amahle avumayo we-serum ezinkomo (n = 420) aqinisekiswa yi-CFT ahlolwe ngokuhlangana namasampula e-serum (n = 446) avela ezinkomeni ezingathelelekile ezahlolwa kuqala ku-RBT, CFT futhi kuqhathaniswa ne-FPA. Inani elinqunyiwe elikhulu elinikezela ukuzwela okuphezulu kokuxilonga (i-Dsn) kanye nokucaciswa kokuxilongwa (i-Dsp) kunqunywe njenge-87 mP kusetshenziswa ukuhlaziywa kwe-ROC. I-Dsn ne-Dsp ye-FPA esebenzisa leli nani elisikiwe libalwe ngama-99.09% - 100% no-68.09% - 76.61% ngokulandelana kwesikhathi sokuzethemba esingu-95% (cl). Indawo engaphansi kwe-Curve noma ijika thizeni (i-AUC) ibalwe ku-0.9842 enephutha elingu-95% elijwayelekile (SE) lika- 0.005532 elinezilinganiso ezinhle nezimbi ze-likehood (+ LR) (-LR) ngo-3.643 no- 1.002, ngokulandelana. I-FPA isebenza njenge-CFT futhi kufanele ibhekwe ngenxa yokunemba eneqiniso kwayo nezinye izinzuzo ezifana nejubane lokuthola imiphumela kanye nenhloso yokuchazwa kwemiphumela engatholakala ngomshini wekhomphuyitha (PHERAstar), i-FPA kufanele ifakwe ekuhlolweni okuvamile ngokujwayelekile kwe-serological ye-brucellosis.