Objective: To investigate the diagnostic value of endoscopic ultrasound-guided fine-needle aspiration (EUS-FNA) for pancreatic lesions. Methods: Forty-five patients diagnosed with pancreatic space occupying lesions in Endoscopy Center of Nantong Tumor Hospital from December 2016 to November 2019 were enrolled. Fine-needle aspiration biopsy was performed under the guidance of endoscopic ultrasonography. The EUS-FNA histological results were compared with the final clinical diagnosis. Results: The sensitivity, specificity and positive predictive values of EUS-FNA for diagnosing pancreatic lessons were 85.71%, 100% and 100%, respectively. The diagnostic accuracy of EUS-FNA was 80%. There was no statistical significance in diagnostic accuracy between the EUS-FNA dignosis and the final diagnosis. The incidence of postoperative complications was 4.44%. There were no serious complications associated with EUS-FNA, such as bleeding, perforation and severe pancreatitis. Conclusion: EUS-FNA is a safe and effective method for the diagnosis of pancreatic space-occupying lesions.