The invention provides an SPR (surface plasma resonance) sensor for detecting HIV (human immunodeficiency virus) related genes and preparation and application thereof; the preparation of the SPR sensor comprises: preparing an entropy-driven strand displacement amplifying system, including, (a), preparing a probe, to be specific, using DNA single strands P, R and Q to prepare three-strand composite PRQ, (b), taking the three-strand composite PRQ obtained in step (a), mixing hybridization solution, the three-strand composite PRQ, single strand F and target sequence T, and hatching to obtain a reaction liquid, (c), fixing a capture probe to the surface of a working chip, inserting the working chip in a surface plasma resonance apparatus, and injecting the reaction liquid of step (b) as a sample to the surface of the working chip wherein the capture probe is fixed so as to obtain the entropy-driven strand displacement amplifying system. The surface plasma resonance sensor and detection system suitable for detecting HIV related genes are successfully constructed herein; by using the sensor of the invention to measure HIV related gears, good sensitivity, stability and reproducibility are achieved.