The invention provides a reagent pre-packaged structure of a centrifugal type micro-fluidic chip as well as a manufacturing method and an application method of the reagent pre-packaged structure, and relates to a manufacturing method and an application method of a reagent pre-packaged structure of a hard polymer centrifugal type micro-fluidic chip. The reagent pre-packaged structure can be used for realizing the pre-packaging and long-term storage of a reagent on the micro-fluidic chip, avoiding the time-consuming manual sample injection or the requirement for a large-sized sample injection device before the chip is used, and ensuring the requirements of practical situations on chip stability and vibration resistance. A laser machining technology is used for machining; an adopted material is PMMA (polymethyl methacrylate); the reagent packaging is realized through a sealing cavity formed by splicing a pressure-sensitive adhesive and PMMA; the starting of prepackaging is realized through the installation of the chip on a centrifugal machine and centrifugal force provided by the centrifugal machine; and the prepackaging and long-term storage requirements of the reagent are effectively satisfied. The reagent pre-packaged structure has the advantages of simple process and high reliability, and is easy to realize and suitable for batch production lines.