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Background and Aim: Endoscopic ultrasound‐guided fine‐needle biopsy (EUS‐FNB) is used to diagnose lesions within or adjacent to the digestive tract. However, there is no report on the overall diagnostic accuracy, technical success, and adverse events of FNB. The aims of this study were to conduct a systematic review and meta‐analysis to comprehensively assess the diagnostic accuracy, technical success, and adverse events of FNB. Methods: Pubmed, Embase, and Cochrane Library databases were searched for relevant articles published in English from January 1998 to May 2019 (No. CRD42019141647). Primary outcomes were EUS‐FNB related diagnostic accuracy rate, technical success rate, and adverse event rate. Results: A total of 51 articles including 5330 patients met our criteria. The overall EUS‐FNB related diagnostic accuracy rate, technical success rate, and adverse event rate was 90.82% [95% confidence interval (CI) 88.69–92.76%], 99.71% [95% CI 99.35–99.93%], and 0.59% [95% CI 0.29–1.0%], respectively. Biopsy with 22G needle could increase the diagnostic accuracy rate and technical success rate to 92.17% [95% CI 89.32–94.61%] and 99.88% [95% CI 99.64–99.99%], respectively, and decrease the adverse event to 0.37% [95% CI 0.08–0.87%]. Moreover, it showed that 22G needle was an independent factor associated with a higher diagnostic accuracy rate and technical success rate and a lower adverse event rate (P = 0.04, P < 0.001, and P = 0.04, respectively) by univariate and multivariate meta‐regression analyses. Conclusion: Endoscopic ultrasound‐guided fine‐needle biopsy is a feasible and safe procedure for lesions within or adjacent to the digestive tract. Biopsy using 22G needle could increase the diagnostic accuracy rate and technical success rate and decrease adverse event rate during the FNB procedure. [ABSTRACT FROM AUTHOR]