Abstract:
Objective: In this study, a paper-based colorimetric sensor was constructed and applied to detect the specific immunoglobulin E (sIgE) of shrimp allergic patients. Methods: The specific antibody (sAb) of the important allergen components in
Litopenaeus vannamei, including tropomyosin (Lit v 1), arginine kinase (Lit v 2) and sarcoplasmic calcium-binding protein (Lit v 4), were prepared and their titer were evaluated. The paper-based colorimetric sensor was prepared with allergens as recognition elements, comprising a covering layer, a working layer and a waste liquid outflow layer. The reaction conditions of the paper-based sensor were optimized and its analytical performance was systematically evaluated. Results: Under the optimal conditions (secondary antibody dilution of 1:2000, target incubation time of 15 min etc.), Lit v 1-sAb, Lit v 2-sAb, Lit v 4-sAb were detected. Theirs recoveries were from 95.14% to 103.25%, with the relative standard deviations of less than 10%. The sensor was applied to the detection of sIgE in serum samples from shrimp allergic patients with a detection limit of 0.22 ng/mL and a quantitation limit of 0.22 ng/mL . In addition, the sensitivities of the Lit v 1-sIgE, Lit v 2-sIgE, and Lit v 4-sIgE ranged from 80.00% to 100%, and the specificities of the three ranged from 91.67% to 100.00%. The consistency of this method with ELISA results reached 88.10%. Conclusion: The sensor has good sensitivity and accuracy. The method is simple, rapid, and suitable for quantitative analysis of allergen component sIgE in shrimp allergic patients. This provided technical support for the component resolved diagnosis of crustacean aquatic products.