A kit based on three-phase enzyme-linked immunoassay and application thereof

CN117491622BActive Publication Date: 2026-07-07SUZHOU UNIV
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Patent Information

Authority / Receiving Office
CN · China
Patent Type
Patents(China)
Current Assignee / Owner
SUZHOU UNIV
Filing Date
2023-10-23
Publication Date
2026-07-07

AI Technical Summary

Technical Problem

Existing 5-fluorouracil detection technologies suffer from inaccuracy, insensitivity, and a tendency to produce false positives. Traditional solid/liquid two-phase interface biosensors exhibit low and fluctuating oxygen concentrations, affecting the accuracy and linearity of the detection.

Method used

A porous hydrophobic composite material was used as the sensor substrate to construct a solid/liquid/gas three-phase interface. Hydrogen peroxide was generated by oxidase catalysis of the substrate, and then reacted with TMB under the catalysis of horseradish peroxidase. This ensured sufficient and stable oxygen supply, thereby improving the enzyme reaction rate and detection performance.

Benefits of technology

It achieves efficient, rapid, and accurate detection of 5-fluorouracil, avoids false positive results, improves detection sensitivity and linear range, reduces detection costs, is easy to operate, and is suitable for widespread application.

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Abstract

This invention provides a kit based on a three-phase enzyme-linked immunosorbent assay (ELISA) and its application. The construction of the three-phase ELISA sensor in the kit mainly includes the following steps: first, selecting a hydrophobic substrate, washing and pretreating it; second, immobilizing the coated antigen and oxidase on the substrate. This invention develops a competitive immunoassay method based on a three-phase substrate and applies it to the detection of 5-fluorouracil. This invention utilizes the specific immunoreaction between a collaboratively developed 5-fluorouracil monoclonal antibody and the antigen to quantitatively detect 5-fluorouracil, improving the selectivity for the target analyte. This immunosensor uses inexpensive equipment, is easy to operate, has high detection efficiency, simple sample processing, and exhibits a wide linear range and a high detection upper limit, making it easy to popularize and cost-effective, thus possessing high practical value.
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