The invention relates to the field of biotechnology and provides a multiple PCR detection primer group, a kit and a detection method for fungus producing aflatoxin. According to the invention, through combined utilization of a fungus universal molecular marker sequence (internal transcribed spacer, ITS) and three key genes, comprising an aflatoxin production regulating gene (aflR), a sterigmatocystin-o-methoxy transferase gene (omt-1), and versicolorin A dehydrogenase gene (ver-1), in the biochemically synthetic process of the aflatoxin, the multiple PCR detection primer group, the kit and the detection method have the technical effects of wide coverage range, strong specificity and high sensitivity during the detection of the fungus producing aflatoxin. The primer group, the kit and the detection method are short in detection time, are simple in operation and are low in device requirement, can save large amount of manpower, materials and costs, are suitable for quick detection and are easy to promote in practical production.