Paper-based ELISA Diagnosis Technology for Human Brucellosis Based on a Multiepitope Fusion Protein

Abstract

Background: At present, as a serious zoonotic infectious disease, the incidence of brucellosis is increasing each year worldwide, exhibiting signs of resurgence. Brucellosis seriously threatens the health of humans, and it is necessary to strengthen the methods utilized for its rapid and accurate diagnosis.Methods: Bioinformatic technology was used to predict B-cell epitopes of the main outer membrane proteins of Brucella and subsequently verified the antigenicity of these epitopes. Prepared a Brucella multiepitope fusion protein and verified the antigenicity of the protein by indirect ELISA. Whatman filter paper was then modified with nano-zinc oxide to construct a paper-based ELISA (p-ELISA) technology for the diagnosis of brucellosis.Results: A total of 22 linear B cell epitopes were predicted. Each epitope could recognize some brucellosis sera. The constructed multiepitope fusion protein had good antigenicity and significantly reduced cross-reaction compared with LPS. The sensitivity and specificity of the method were 92.38% and 98.35%, the positive predictive value was 98.26%, and the negative predictive value was 91.67%.Conclusions: A multiepitope fusion protein of Brucella was successfully prepared, and a rapid diagnostic technique for brucellosis was established. This technology has potential application value and can be used for the rapid diagnosis of brucellosis.