The present invention discloses a manufacturing method of a test strip, which comprises making a first semi-finished product and a second semi-finished product. The manufacturing process of the first semi-finished product comprises: providing a substrate, forming a plurality of electrodes on the substrate, forming a supporting layer with a plurality of channels on the substrate, and providing a reaction material to fill in the channels. The manufacturing process of the second semi-finished product comprises: providing a lid, forming a hydrophilic layer on a first surface of the lid, forming an adhesive layer on the first surface without the hydrophilic layer. Thereafter, a test strip assembly is formed by adhering the channels of the first semi-finished product to the hydrophilic layer of the second semi-finished product. Finally, a plurality of test strips are produced by cutting the test strip assembly along a first axis of the substrate.