The invention belongs to the technical field of medical examination, and particularly relates to a rapid detection kit for bacterial vaginosis, a preparation method and application thereof. The kit comprises a reagent R1 and a reagent R2, and the reagent R1 is composed of 5-bromine-4-chlorine-3-Indolyl alpha-D-N-acetylneuramidate, polyethylene glycol 200 and phosphate buffer solution; and the reagent R2 is composed of 4-amino antipyrine and the phosphate buffer solution. The rapid detection kit for the bacterial vaginosis has the advantages of high determination sensitivity, high accuracy of detection results, good repeatability and high stability, and is an ideal rapid detection kit for the bacterial vaginosis, and the popularization and application of the kit are facilitated.