Objective Two models of rat kidney stone disease were constructed to provide support for exploration of the etiology of kidney stones. Methods Sixty rats were randomly divided into a nanobacteria group (NB group, tail vein injection of 1. 2 mL NB suspension, 30 rats) and an ethylene glycol group (EG group, 1. 25% ethylene glycol water +1% ammonium chloride 2 mL/ d gavage, gavage duration of 2 w, 30 rats). The experimental period was 10 w. In each group, 3 rats were sacrificed each week, with blood, urine, and kidney specimens being collected before sacrifice. Results There were no significant differences in the general conditions between the two groups, including no difference in renal body ratio ( P > 0. 05). There was also no significant difference in the number of pathological crystallizations. In addition, there were no significant differences of serum calcium, magnesium, phosphorus, creatinine, and uric acid between the two groups ( P > 0. 05), and also no difference of urea ( P > 0. 05). There were no significant differences of urine calcium, urine pH, urine specific gravity, and 24 h urine volume between the two groups ( P > 0. 05). Conclusions No statistically significant differences in terms of research indicators were identified between the two modeling method, indicating the feasibility of nanobacterial modeling. Compared with the conventional method of using ethylene glycol, NB has the advantage of being milder and being derived from kidney stones themselves. This model is more similar to the formation of human kidney stones and more conducive to study of the causes of kidney stones.