The invention discloses a loop-mediated isothermal amplification reagent capable of being transported at a normal temperature as well as a preparation method and application of the reagent. The reagent comprises Bst DNA Polymerase, dNTP, a primer mixture, an indicator, a protective agent and an aerosol pollution preventive. The preparation method comprises the following steps: mixing the previouscomponents, and performing freeze drying at a temperature of 70-80 DEG C below zero for 8-16 hours, thereby obtaining the loop-mediated isothermal amplification reagent. A loop-mediated isothermal amplification reagent used for nucleic acid test is prepared in the invention, can be transported and preserved at the normal temperature, and is convenient to transport and excellent in stability; meanwhile, the problem that popularization of a LAMP (Loop-mediated Isothermal Amplification) technology is limited by aerosol pollution is solved, and the application and popularization difficulty of theLAMP technology is greatly reduced. The activity of the Bst DNA Polymerase in the reagent is 150%, the reagent can be preserved at the normal temperature of one year or longer, only a buffer solutionneeds to be added during use, the operation is simple, the activity of DNA polymerase is high, the detection sensitivity is loss-free, the interference of the protective agent is low, and the cost islow.