Zinc sulfide nano clusters were synthesized and characterized. A kind of method using zinc sulfide nanoparticles cluster cation exchange reaction(CX) to detect trace biological molecules was established. Non fluorescent ZnS nanoparticles (NCCs) were synthesized and characterized. The property of nano clusters directly influences the detection results. Through transmission electron microscopy images and X-ray diffraction, nano clusters which could quickly release a mass of Zn2+ from rapid cation exchange reaction were known to be porous and generate fluorescence signal under the action of zinc reagent. The external crystal arranges loosely compared to the internal, which is conducive to rapid cation exchange, and the crystal size is related to heating time. It was demonstrated that the smallest nanocluster had a relative large surface area and higher cationic exchange efficiency through the determination of the specific surface area of nano clusters for detecting surface area and pore size. Three methods (acid dissolution method, cation exchange and micro wave aided by cation exchange) which effected Zn2+ release performance were experimented. It turned out that microwave auxiliary cation exchange method had high SNR, simple operation, and could be used in zinc sulfide nanoparticle immunoassay. Having compared the relations between the release efficiency, target binding force of Zn2+ and its average diameter, the results show that the nano cluster size of 44 nm exhibits the highest cation exchange efficiency. All these features make the ZnS nanocluster cation exchange amplifier to be a highly sensitive, fairly biocompatible, low-cost and environment friendly detection tool in the detection of biomolecules.