The invention provides a procalcitonin (PCT) determination kit. The kit contains a reagent R1 and a reagent R2, wherein the reagent R1 is prepared from the following components: a piperazine-1, 4-diethylsulfonic acid (PIPES) buffer solution, NaCl, MgCl2, chondroitin sulfate, hyaluronic acid, a surfactant and a preservative. And the reagent R2 is prepared from piperazine-1, 4-diethylsulfonic acid (PIPES) buffer solution, goat anti-human PCT antibody coated latex particles, a suspending aid, a stabilizer and a preservative. The invention further provides a preparation method and application of the kit, the kit can effectively avoid interference of chyle particles, latex particles of different specifications, novel surfactants and different protective agents and suspending agents are adoptedat the same time, and the kit is a liquid kit which is high in stability, high in sensitivity, good in repeatability and low in cost.