The invention relates to a low abundant
protein absolute quantification method based on a digital
immunoassay technology. The method comprises the following steps: carrying out an immunoreaction on captured beads, target antigens and detected particles so as to form an
immune complex, eluting the detected particles in the
immune complex, collecting and concentrating the eluant, transferring the concentrated solution to a micro-fluidic particle counting
chip, depositing and fixing the detected particles on the micro-fluidic particle counting
chip after a
solvent in the concentrated solution iscompletely volatilized, performing imaging recording on all the detected particles fixed on the micro-fluidic particle counting
chip, and counting the quantity of the detected particles on the micro-fluidic particle counting chip, thereby obtaining the molecular number of the
target protein. The method disclosed by the invention has the advantages that ultra-sensitive and
absolute quantification analysis of
disease-related proteins in clinical samples can be realized, any standard substance is not needed, a
standard curve does not need to be drawn, and the method has the advantages of being wide in application range, high in detection sensitivity, high in analytical result accuracy, excellent in precision degree and high in
throughput.