Abstract: Objective:To establish a PCR melting curve method for the identification of Mycobacterium avium (MA) based on SYBR Green Ⅰ fluorescent dye, and then provide a simple, accurate molecular biological method for the clinical identification of MA. Methods:The primers were designed based on the specific insertion sequence IS1311 of MA as the detection target. At the annealing temperature of 60 ℃, a SYBR Green Ⅰ fluorescence PCR melting curve method was established for the identification of MA. The specificity and the detection limit of the established method were evaluated by detecting 21 reference strains of mycobacterium including MA and 5 common pathogens including Staphylococcus aureus. Using the sequencing of PCR products of hsp65 and 16S rDNA as the “gold standard”, 200 clinical isolates of mycobacterium were identified, and then the accuracy of the established method for the identification of MA was evaluated. Results:The established SYBR Green Ⅰ fluorescence PCR melting curve method were able to accurately identify MA in 21 reference strains of mycobacterium and 5 pathogenic bacteria within 30 cycles, and the detection limit was 5.6×10^-2 ng/μL. Among 200 clinical isolates of mycobacterium, 16 strains of MA and 184 strains of other mycobacteria were identified by PCR sequencing, while 15 strains of MA and 185 strains of non-MA were identified by the established SYBR Green Ⅰ fluorescence PCR melting curve method. There was no significant difference in the identification rate of MA between PCR sequencing and the established method in this study (P＞0.05), and there were high coincidence rate (99.5%, 199/200) and high consistency (Kappa value＞0.75, P＜0.01) between them. The sensitivity, specificity, positive predictive value and negative predictive value of the established method were 93.8% (15/16), 100.0% (184/184), 100.0% (15/15) and 99.5% (184/185), respectively. Conclusion:The established SYBR Green Ⅰ fluorescence PCR melting curve method based on the specific repeat sequence IS1311 of MA has high sensitivity and specificity, which provides a new way for the clinical detection of MA.