The invention discloses fluorescent immunochromatography test paper and a preparation method thereof. The test paper is formed by sequentially overlapping a sample pad (1), an
antibody carrying film (2) and absorbent paper (3) through a base plate (4) which is provided with binding agents, the
antibody carrying film (2) is coated with a marker mixture
joint line (5), a testing line (6) and a
quality control line (7) in sequence from the end close to the sample pad (1), and the marker mixture
joint line (5) is formed by arranging a marker mixture obtained by
coupling fluorescent microspheres with an
antibody 1 and
biotin within a pretreatment area (8) which is pretreated with the antibody carrying film in a
coating mode through a crossed film
metal-spraying instrument. By means of the test paper, the problems that the result is not accurate caused by insufficient release amount due to the fact that excessive binding materials remain on glass fibers are solved, the tested
fluorescence value is raised, and the sensitivity or detection upper limit is lifted. Meanwhile, in the production, the antibody carrying film is coated with the marker mixture obtained by
coupling the
fluorescent microspheres with the antibody 1 and the
biotin, the marker mixture
joint line, the T line and the C line can be coated simultaneously, time and processes are saved, and the production efficiency is improved.