The guideline for the diagnosis and treatment of monkeypox (2022 edition) issued by National Health Commission of the People's Republic of China introduces the key knowledge of the diagnosis and treatment of human monkeypox (HMPX) and does not systematically introduce the sampling methods and requirements of specimens for HMPX etiology testing and the discrepancy in diagnostic criteria between China and overseas. However, the doctors who are not engaged in dermatology lack understanding of the sampling methods and requirements of specimens for laboratory diagnosis of HMPX, and there are few relevant references available. This article collects the information on the diagnosis and treatment of HMPX, so as to provide a reference for learning, understanding, and application of this guideline.