In recent years, endoscopic ultrasound-guided fine-needle aspiration/biopsy (EUS-FNA/B) has been widely applied throughout China and plays an increasingly important role in the diagnosis and treatment of diseases of digestive tract or adjacent organs. With the development and innovation of perceptions and techniques, the standardization in clinical practice of EUS-FNA/B becomes an urgent need. The guideline presented herein is based on existing guidelines and makes substantial updates on frontier progress in this field for the past few years. Based on the most current evidence, a total of 21 clinical concerns and 37 recommendations are proposed including the indications and contraindications, skills and techniques, specimen processing, perioperative management, and training and education of EUS-FNA/B. It is expected to guide the clinical application of EUS-FNA/B.