Milk Non-Enzymatic Immunofiltration Assay "mnerifa": An Alternative Rapid Bovine Milk Test for Anti-Brucella Antibody Detection

Abstract

Rapid milk tests for the indirect diagnosis of brucellosis are limited and generally not preferred due to insufficient diagnostic performances and inappropriate handling process. In this study, a rapid immunofiltration assay known as Non-Enzymatic Immunofiltration Assay (NERIFA) was evaluated based on diagnostic performances and applicability for the detection of anti-Brucella antibodies in bovine milk samples as milk NERIFA (mNERIFA). For this purpose, wheys, that were obtained from rennin precipitated milk constituted with reference sera, were used as test material. Besides, an enzyme-linked immunosorbent assay (ELISA) was also developed for the evaluation of mNERIFA. After approval, the ELISA was evaluated as important (P < 0.001) by Receiver Operating Procedures (ROC) analysis based on the diagnostic index (J) score specified as the Youden index (J: 0.963) and the Area Under the Curve (AUC) value (0.996), its results compared with mNERIFA by kappa statistical analysis. ELISA's comparison with mNERIFA based on diagnostic performances indicated that ELISA's performance was in perfect agreement with mNERIFA (.=0.97) and the agreement was 0.97% (0.92-0.99%) with CI 95%. In the optimization process of mNERIFA, it was found that there was no significant difference between the individual and pooled whey samples (P > 0.05), and a pool of 5 samples can be used instead of individual testing. In this study, it was concluded that mNERIFA may be recommended as a rapid test for anti-Brucella antibody detection in bovine milk samples based on the diagnostic performances, applicability, and pooling capacity.