The invention provides a low-serum efficient culture medium for culturing mycoplasma hyopneumoniae and a preparation method thereof. The low-serum efficient culture medium comprises a base medium and an addition bag. The base medium comprises M199, brain extract powder, lactoalbumin hydrolysate, BSA, glucose, fresh yeast extract, Hepes, iron citrate, calcium chloride, 2H2O, phenol red and dd H2O. The addition bag comprises cholesterol, linoleic acid, linolenic acid, palmitic acid, ethanol and porcine serum. Through use of some hydrolyzates and lipid, a serum addition ratio is less than 5% so that a cost is reduced, the quality of mycoplasma is improved, and mycoplasma hyopneumoniae antigen titer is improved.