Immunoassay can detect trace substances in body fluids. Immune solid phase carrier is a surface reaction platform for the specific binding of antigen and antibody in immunoassay detection technology. Its performance directly determines the sensitivity and specificity of immunoassay. In this project, the specially modified nuclear track etched membrane is used as the immune solid-phase carrier. Using the characteristics of accurate and controllable pore diameter, a large number of micron through channels and smooth surface of dielectric film, supplemented with optimized liquid channel design, an immunoassay detection technology platform is developed, which has the obvious advantages of high sensitivity, strong anti-interference ability and high throughput.
This project innovatively proposes to develop two sets of membrane surface and pore wall modification processes with nuclear track etched membrane as immune solid carrier; The antibody coating process was systematically studied and determined; Time resolved fluorescence immunoassay (TRFIA-NTM) and enzyme linked immunosorbent assay (ELISA-NTM) based on grafted nuclear track etched membrane were established, and the corresponding detection reagent card was developed. For the detection of SARS-CoV-2 nucleocapsid protein, the detection sensitivity of TRFIA-NTM can reach 0.3pg/ml, which is two orders of magnitude higher than that under NC membrane, and the detection sensitivity is significantly improved. The linear range is 1 ~ 80ng / ml, and the detection time at room temperature is only 15min; ELISA-NTM detection sensitivity reached 0.1pg / ml, and the detection time is 45min at room temperature.
This method has the advantages of high sensitivity, high throughput, low background, rapid response and simple operation. It is an original scientific and technological achievement with independent intellectual property rights. It can be easily used to build up a full-automatic, rapid and large sample size detection platform, and has possibility to develop into a singl.
