Objective To investigate the clinical significance of serum myoglobin (Mb), cardiac troponin T(cTnT) and high-sensitivity C-reactive protein (hs-CRP) levels in patients with congestive heart failure. Methods 120 patients with congestive heart failure in the hospital were selected as the observation group, and 120 healthy people were used as the control group at the same time. Serum Mb, cTnT and hsCRP were detected. The diagnostic efficiencies of Mb, cTnT and hs-CRP for patients with congestive heart failure were evaluated. Serum Mb, cTnT and hsCRP levels in patients with different NYHA grades of congestive heart failure were compared. Results The levels of serum Mb, cTnT and hs-CRP in the observation group were significantly higher than those in the control group (P＜0.05). The sensitivities of serum Mb, cTnT and hs-CRP for diagnosis of congestive heart failure were 91.67%, 70.83% and 82.50%, respectively.The specificities were 78.33%, 83.33% and 63.33%, respectively. The sensitivity, specificity, accuracy, positive predictive value and negative predictive value of combined diagnosis of serum Mb, cTnT and hs-CRP for congestive heart failure were higher than those of single detection. Serum Mb, cTnT and hs-CRP levels were significantly higher in NYHA grade IV patients than in NYHA grade Ⅱ and grade Ⅲ patients (P＜0.05). Serum Mb, cTnT and hs-CRP levels in NYHA grade Ⅲ patients were significantly higher than those in NYHA grade Ⅱ patients (P＜0.05). Conclusion Combined detection of serum Mb, cTnT and hs-CRP can improve the diagnostic efficiency of congestive heart failure. Besides, serum Mb, cTnT and hs-CRP levels can be used to effectively assess the NYHA grade, can be used as an index to evaluate the severity of congestive heart failure.