The invention relates to a Toxoplasma gondii rapid detection kit and a preparation method thereof. The kit is based on the molecular biology and the nucleoside triphosphate hydrolase (NTPase) gene with high conservation of Toxoplasma gondii to adopt the loop-mediated isothermal amplification (LAMP) method to perform the rapid detection of Toxoplasma gondii. The kit contains LAMP reaction solution, primer, Bst DNA polymerase, color reagent, standard positive template and dd H2O. The kit of the invention is characterized by high sensitivity and specificity, rapid reaction, simple operation and the like, does not require special instrument and equipment and solves the defects of the prior art such as long detection time of toxoplasmosis, large workload and complicated operation. The kit of the invention can be used to detect whether a clinical sample contains Toxoplasma gondii and used in the early diagnosis and the molecular epidemiological study of toxoplasmosis.