Digital immunoassay detection and analysis method for precise quantification of proteins

A digital immunoassay device for quantification of proteins, wherein the device comprises:

• (a) a mixing module, which is configured to form an aqueous phase mixture comprising the following components:
• (A) a positive signal complex:
• a magnetic bead—antibody—antigen—detection antibody—enzyme label, and/or magnetic bead—antibody—antigen—detection antibody complex;
• and
• (B) a negative signal complex:
• a magnetic bead—antibody complex;
• (b) a shearing module, which is configured to add a chromogenic substrate and an oil phase to the aqueous phase mixture to form a first mixture;
• and perform a mechanical shearing to form a sheared water-in-oil second mixture, which contains first and second micro droplets;
• the first micro droplet is a micro droplet containing a positive signal complex;
• the second micro droplet is a micro droplet containing a negative signal complex;
• and
• (c) a digital detection module, which is configured to measure the first micro droplet, thereby obtaining the number of the first micro droplet and obtaining the quantitative detection result of the proteins.