Objective To establish a visual loop-mediated isothermal amplification detection method for rapid detection of Staphylococcus epidermidis. Methods A loop-mediated isothermal amplification system was established by selecting specific primers for SesB gene of Staphylococcus epidermidis. The visualization method of the reaction, the concentration of MgSO ₄, betaine in the system and the reaction temperature, time were optimized. The sensitivity and specificity of the optimized LAMP reaction and PCR reaction were compared. Results The optimal MgSO ₄ concentration in the loop-mediated isothermal amplification reaction system was 4mM. The optimal betaine concentration was 0.4M. The optimal reaction temperature and time were 63℃ for 60 min. The most suitable visualization method was calcein chromogenic method, and the visualization results were basically consistent with the results of agarose gel electrophoresis. Both LAMP and PCR reactions had good specificity. The sensitivity of LAMP reaction was 10 copies/μL, and the sensitivity of PCR reaction was 100 copies/μL. Conclusion LAMP assay is superior to PCR in the detection of Staphylococcus epidermidis infection. It is more suitable for grass-roots and remote areas due to its simple operation and short reaction time.