The invention discloses a soil toxoplasma gondii detection kit and a preparation method thereof. The soil toxoplasma gondii detection kit comprises a 10*PCR (Polymerase Chain Reaction) liquid, a primer mixture, a thermophilic DNA (Deoxyribonucleic Acid) polymerase, a bovine serum albumin and a standard male template. The kit is characterized in that the primer mixture comprises 0.2pmol of an upstream primer TX1, 10pmol of a middle primer TX2 and 10pmol of a downstream primer TX3. The kit disclosed by the invention is established on the basis of molecular biology, is based on high-conservation 529bp repetitive sequence genes of toxoplasma gondii and detects the soil toxoplasma gondii by applying a half-set type PCR method; the kit has the characteristics of high sensitivity, strong specificity, simplicity in operation and the like; the defects that the relevance ratio of detection on toxoplasma gondii oocysts in the soil is low, the working amount is great, the operation is complicated, a diagnostic technique is insufficient and the like can be solved; and the soil toxoplasma gondii detection kit can be used for detecting whether a soil sample contains the toxoplasma gondii oocysts or not, can be used for carrying out molecular epidemiological investigation on the soil toxoplasma gondii and can be used for monitoring an ambient environment in real time.