Introduction: Serum biomarkers are useful for physicians to screen, diagnose, and treat lung cancer. However, the diagnostic value of a single marker is limited by its sensitivity and specificity. Therefore, combination marker panel is frequently chosen in the clinic. However, combination panel has increased costs. China is a developing country with a large population where nearly 70% of the people live in poor rural areas. Objective To evaluate the efficiency of 5 tumor biomarkers, namely carbohydrate antigen-125(CA-125), carbohydrate antigen-199(CA-199), carcino embryonic antigen(CEA), neuron-specific enolase (NSE), and Cytokeratin 19(CYFRA21-1), independently or in combination for lung cancer diagnosis, pathological type and clinical stage. Methods: This is a prospective study of sixteen months done in the Respiratory Department of Medicine, Zhengzhou University, China from September 2015 to December 2016. Electrochemiluminescence immunization was used to determine biomarker levels expressed in 259 patients with lung cancer and 50 benign lesion controls. Results: The observed levels of CA125, CA199, CEA, NSE and CYFRA 21-1 in patients with pathologically confirmed lung cancer were significantly higher than those in patients with benign pulmonary diseases. Adenocarcinoma, squamous cell carcinoma, and small cell lung carcinoma of the lung were associated with the highest observed levels of CEA, CYFRA21-1, and NSE, respectively. The combination of 4 tumor biomarkers, namely, CAl25+CEA+NSE+CYFRA2l-1 showed 85.63% sensitivity for lung cancer. The ROC of CYFRA2l-1 was 0.840 ± 0.032(95%CI,0.777-0.904). The positive rates and level of CA125, CEA, CYFRA21-1 is different in clinical stages of lung cancer. Conclusion: CA125, CA199, CEA, NSE and CYFRA 21-1 are valuable in the diagnosis, pathological type and clinical stage of lung cancer, especially CYFRA 21-1. The combination of CAl25+CEA+NSE+CYFRA2l-1 showed higher sensitivity for lung cancer. CA125, CEA, CYFRA21-1 play an important role in clinical stage. Funding Statement: None. Declaration of Interests: None. Ethics Approval Statement: The study was approved by the hospital ethical committee, and written consent was obtained from each subject for their participation in the study.