Objective: To analyze the testing value of hierarchical testing methods in clinical blood lipid biochemical testing. Methods: 120 subjects who required blood lipid biochemical testing received by our hospital from January 2024 to January 2025 were selected and divided into a hierarchical testing group (60 cases) and a routine testing group (60 cases) according to different testing methods. The routine inspection group adopts the full-item routine inspection method, and the graded inspection group adopts the graded inspection method, that is, the basic items of total cholesterol (TC) and triglycerides (TG) are first detected, and then high-density lipoprotein cholesterol (HDL-C), low-density lipoprotein cholesterol (LDL-C) and other items are selectively tested based on the results of the basic items. Compare the accuracy of test results, test time, test costs, and subject satisfaction between the two groups. Results: When comparing the test results of TC, TG, HDL-C, and LDL-C between the two groups, the difference was not statistically significant (p > 0.05); the inspection time of the graded inspection group was shorter than that of the conventional inspection group, the inspection cost was lower than that of the conventional inspection group, and the subject satisfaction was higher than that of the conventional inspection group (p < 0.05). Conclusion: The use of hierarchical testing methods in clinical blood lipid biochemical testing can shorten testing time, reduce testing costs, and improve subject satisfaction while ensuring testing accuracy, and has high clinical application value.