Objective To evaluate the diagnostic value of contrast-enhanced ultrasound-guided percutaneous liver biopsy for focal liver lesions. Methods 354 patients with 401 lesions were retrospectively analyzed from May 2015 to may 2019 in Gulou hospital. 354 patients with 401 lesions were divided into two groups: Group A (contrast-enhanced ultrasound-guided percutaneous liver puncture, n = 65 patients with 77 lesions ) and Group B (ultrasound-guided percutaneous liver puncture, n = 289 patients with 324 lesions). The difference in diagnostic accuracy between the two groups was analyzed to evaluate the value of performing contrast-enhanced sonography before biopsy. Results The overall diagnostic accuracy of contrast-enhanced ultrasound-guided percutaneous liver puncture group was better than that of ultrasound-guided percutaneous liver puncture group (95.1% vs 85.7%, P = 0.003), and the diagnostic accuracy of malignant lesions was better than that of ultrasound-guided percutaneous liver puncture group (95.3% vs 84.7%, P = 0.008). The specificity and positive predictive value of contrast-enhanced ultrasound-guided percutaneous liver puncture group were higher than those of ultrasound-guided percutaneous group (83.8% and 64.0%, P = 0.036; 95.3% vs 84.7%, P = 0.008). Especially for lesions less than 2 cm, the diagnostic accuracy of contrast-enhanced ultrasound-guided percutaneous liver puncture group (96.3%, 52 / 54) was significantly better than that of ultrasound-guided percutaneous liver puncture (74.1%, 20 / 270), X2 = 6.891, P = 0.009. Conclusions Contrast-enhanced ultrasound-guided percutaneous liver biopsy for focal liver lesions is easy to operate with a low risk, and can significantly improve the diagnostic raccuracy, especially for lesions less than 2 cm in size.