An innovative lateral flow sulphur autotrophic denitrification (LFSAD) reactor was developed in this study; the treatment performance was evaluated and compared with traditional sulphur/limestone autotrophic denitrification (SLAD) reactor. Results showed that nitrite accumulation in the LFSAD reactor was less than 1.0 mg/L during the whole operation. Denitrification rate increased with the increased initial alkalinity and was approaching saturation when initial alkalinity exceeded 2.5 times the theoretical value. Higher influent nitrate concentration could facilitate nitrate removal capacity. In addition, denitrification efficiency could be promoted under an appropriate reflux ratio, and the highest nitrate removal percentage was achieved under reflux ratio of 200%, increased by 23.8% than that without reflux. Running resistance was only about 1/9 of that in SLAD reactor with equal amount of nitrate removed, which was the prominent excellence of the new reactor. In short, this study indicated that the developed reactor was feasible for nitrate removal from waters with lower concentrations, including contaminated surface water, groundwater or secondary effluent of municipal wastewater treatment with fairly low running resistance. The innovation in reactor design in this study may bring forth new ideas of reactor development of sulphur autotrophic denitrification for nitrate-contaminated water treatment.