The invention discloses a penitrem A combined antigen as well as preparation and an application of an antibody of the antigen. The median inhibitory dose (IC50), detected by a penitrem A enzyme-linked immunosorbent assay kit, of penitrem A for a reaction between a specific antibody of the penitrem A and penitrem A coating antigens is 100 ng / mL, the lowest detection limit of penitrem A is 30 ng / mL, and the cross reaction rate of penitrem A and an aflatoxin B1 sample, the cross reaction rate of penitrem A and a cyclopiazonic acid sample, the cross reaction rate of penitrem A and a vomitoxin and the cross reaction rate of penitrem A and a citreoviridin sample are all smaller than 0.1%. The specific antibody of penitrem A in the penitrem A enzyme-linked immunosorbent assay kit has the characteristics of high potency, high sensitivity and high specificity and can be used for quickly, sensitively and easily detecting the residual amount of penitrem A in animal-origin food.