The invention is applicable to the technical field of in-vitro diagnosis and immunodetection, and provides a 25-hydroxyvitamin D dissociation-
neutralization-quantification integrated micro-fluidic
chip which comprises a closed shell, a PET (
polyethylene terephthalate) film layer and a PMMA (
polymethyl methacrylate) substrate layer which are sequentially assembled from top to bottom, the PET film layer is provided with a continuous micro-
channel network, the micro-
channel network is formed by alternately connecting straight channels and serpentine channels in series, and an alkalization dissociation area, an acidification
neutralization area, a
delay diffusion area and a detection area are sequentially integrated in the liquid flow direction; according to the invention, the
chip is self-driven by capillary force and is combined with the mixing enhancement and driving force supplement effects of the micro-column array, so that efficient dissociation, rapid
neutralization and sequential competitive immunodetection of 25-hydroxyvitamin D in a sample are realized, an external pump valve or artificial
liquid transfer is not needed, and full-automatic and rapid quantitative detection of'sample input-result output 'is achieved; the problems of complicated operation, difficulty in
miniaturization and large detection error in the prior art are solved, and the method is suitable for real-time detection scenes.