Expression and serodiagnostic efficacy of a novel echinococcosis-specific recombinant fusion antigen rAgB8/1-Em18-Eg95.

Abstract

Echinococcosis lacks sensitive serological diagnostic tools. The echinococcosis-specific antigens Eg95, AgB8/1 and the Em18 gene sequences were fused and expressed as the novel recombinant antigens rAgB8/1-Em18-Eg95 (T3) and rEm18-Eg95 (T2), used for the diagnosis of hydatid disease, prepared into an enzyme-linked immunosorbent reaction (ELISA) kit, and evaluated for their serological diagnostic value. The relative molecular weight of the T3 protein was 88.1 kDa, the purified concentration was 1.5 mg mL⁻¹, and the purity was 80%. The relative molecular weight of T2 protein was 79.9 kDa, the total protein concentration was 0.5 mg mL⁻¹, and the purity was less than 50%. The overall coincidence rate of T2 protein was low, and it was impossible to distinguish between negative and positive sera. The T3 antigen was coated at 1.0 μg mL⁻¹, the cutoff value was 0.5271, and the serum dilution ratio was 1:400. A T3 ELISA kits (96 tests) was constructed to detect the serum of 272 clinically and pathologically confirmed cases. The sensitivity of T3 was 93.8%, and the specificity was 83.3%. The parasite cross-reaction was 30%. Satisfactorily, the Pearson correlation coefficient between the T3 OD value and lesion diameter was 0.707, showing a strong correlation. T3 exhibits better antigenicity than T2, and the prepared T3 ELISA diagnostic kits reached the laboratory diagnostic level of a commercial kits. T3 can distinguish human cystic echinococcosis (CE) and alveolar echinococcosis (AE) more significantly and predict the diameter of lesions according to the OD value, which provides practical value for drug or surgical efficacy.