Abstract: Objective：To establish a method of loopmediated isothermal amplification assay (LAMP) for rapid detecting Staphylococcus aureus (S.aureus) in clinical samples. Methods：A highly specific set of four primers, two inner primers (the forward inner primer termed FIP and the backward inner primer termed BIP) and two outer primers (F3 and B3) that recognize a total of six distinct sequences at target DNA, was designed to detect femA gene ofS.aureus and LAMP forS.aureus was developed. Forty clinical isolates of S.aureus, 12 other grampositive cocci and 8 kinds of gramnegative bacteria was detected, and the specificity of the method was evaluated. The standard stain ofS.aureus ATCC 25923 was diluted by 10ⁿ, and the sensitivity was evaluated. S.aureus in 60 clinical throat swab samples was detected by LAMP and culture method was performed respectively. Results：The optimal incubation temperature of LAMP was 65 ℃. The 40 clinical isolates of S.aureus were detected by LAMP and the positive rate was 100%. No specific reactions were found in other bacteria stains. The LAMP assay showed a limit of 14 CFU/test. The sensitivity，specificity，positive predictive value and negative predictive value of LAMP by detecting sixty throat swab samples was 87.5%，100%，100% and 98.1%, respectively. Conclusion：The developed LAMP is very simple, rapid, sensitive and specific method and is available for rapid identification for clinic swab specimen within 60 min.