ArticleMikrochimica acta2026
An ultrasensitive double-antigen sandwich time-resolved fluorescent immunochromatographic assay for quantification of Brucella antibodies.
Article in Mikrochimica acta, 2026. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Not yet cited in PubMed.
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Abstract
Brucellosis is a zoonotic disease that poses a significant threat to public health and the development of animal husbandry, making early and rapid diagnosis essential for effective control. In this study, a double-antigen sandwich time-resolved fluorescent immunochromatographic assay (EuCM-TRFICA) utilizing europium chelate microsphere (EuCM) was developed for the highly sensitive quantification of Brucella antibodies in human serum. This is the first report of a double-antigen sandwich TRFICA utilizing Brucella LPS, which enables the detection of total antibodies with superior sensitivity compared to the previously reported colloidal gold immunochromatographic assay (AuNP-ICA) and time-resolved fluorescence LFIA (TF-LFIA). Under the optimal parameters, EuCM-TRFICA exhibited a satisfactory linear range from 0.156 to 5.00 IU/mL, and its limit of detection (0.145 IU/mL) was lower than that (0.3125 IU/mL) of AuNP-ICA. Furthermore, it effectively identified positive samples missed by the serum agglutination test (SAT) or the rose bengal test (RBT). EuCM-TRFICA exhibited high specificity, with only weak cross-reactivity observed against Yersinia enterocolitica O9, which has been reported to share similar antigenic epitopes. Precision evaluation showed that both intra- and inter-assay coefficients of variation (CV) were below 10.55%. The recoveries of spiked samples ranged from 91.94% to 108.85%, confirming reliable accuracy. In the testing of 49 confirmed positive samples and 60 negative samples, EuCM-TRFICA achieved 100% accuracy, outperforming SAT (92.7%), RBT (86.2%), and ELISA (97.2%). In conclusion, the EuCM-TRFICA exhibits excellent performance in terms of sensitivity, quantitative capability, and detection speed. It features simple operation, a short detection time of 15 min, quantitative readout, and controllable costs. This assay shows promise as a potentially efficient technical tool for early on-site screening and epidemic surveillance in this preliminary cohort, which may be valuable for managing human brucellosis.
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