The invention discloses an immune chromatography test paper strip based on an up-conversion luminescence technology. The up-conversion luminescence material, i.e. UCP, is used as a biological marker, and the result can be presented in a visible light signal form under the radiation of infrared light and can be interpreted by an instrument, thereby quantitative detection on the target detected object is realized. The test paper strip mainly comprises a sample pad, a binding pad or a binder release pad, an analyzing membrane, a water absorbing pad and a plastic back plate. The invention also discloses a method for preparing the test paper strip, and application in the biological sample quantitative detection. According to different immune reaction modes of the substances to be detected, the test paper strip is classified according to a sandwich mode, a competition mode and an indirection mode, and various modes of the test paper strips can carry out quick and sensitive qualitative and quantitative detection and analysis on different substances to be detected in the sample.