Application value of hypersensitive C-reactive protein (hs-CRP), blood lactic acid (LAC) and myoglobin (Mb) in the combined detection of myocarditis was explored. A total of 107 patients with suspected myocarditis treated in The Second Nanning People's Hospital from January 2015 to December 2017 were retrospectively analyzed, of whom 81 patients diagnosed with myocarditis were enrolled into the research group, and 26 non-myocarditis individuals were enrolled into the control group. Fasting venous blood was drawn from all patients to detect the hs-CRP, LAC and Mb, and their levels were compared and analyzed between the two groups. Moreover, the coincidence rate, sensitivity and specificity of single detection and combined detection by hs-CRP, LAC and Mb in the diagnosis of myocarditis were compared and analyzed. There were no significant differences in the sex, age, smoking status, alcohol consumption, chest distress, palpitation, angina and dyspnea between the two groups (P>0.05), proving that patients in both groups were comparable. The levels of hs-CRP, LAC and Mb in the research group were significantly higher than those in the control group, displaying statistically significant differences (P<0.05). According to the receiver operating characteristic (ROC) curve, the area under the curve (AUC), coincidence rate, sensitivity and specificity in the diagnosis of myocarditis, respectively, were 0.610 (95% CI: 0.495–0.725), 58.88, 51.85 and 80.77% for hs-CRP, 0.657 (95% CI: 0.551–0.763), 58.88, 54.32 and 73.08% for LAC, 0.747 (95% CI: 0.651–0.843), 69.16, 64.20 and 84.62% for Mb, and 0.773 (95% CI: 0.680–0.867), 76.64, 79.01 and 69.23% for combined detection. Hs-CRP, LAC and Mb are highly expressed in the serum of patients with myocarditis, and their combined detection has guiding significance for the prevention and treatment of myocarditis.