Objective To understand the clinical features and epidemiological characteristics of monkeypox cases in Shangcheng district, Hangzhou city, providing reference basis for monkeypox epidemic prevention and control work. Methods Data on 35 monkeypox cases detected by medical institutions in Shangcheng district, Hangzhou city from July to December 2023 were collected, including demographic information, clinical manifestations, epidemiological information, laboratory test results, and the clinical features and epidemiological characteristics of the cases were analyzed. Results All 35 monkeypox cases were male sexual contacts, with a median age of 28 years (range: 20 to 37 years). None had a history of smallpox vaccination, and all of them were identified through active medical consultation. Among them, 25 cases (71.43%) admitted to having had male homosexual contact within 21 days before onset, with an interval from last exposure to onset ranging from 1 to 14 days, median (Q1, Q3) of 6 (5, 9) days. Clinical symptoms mainly included fever, rash, inguinal lymph node pain, and muscle soreness; 24 cases (68.57%) presented with fever, all 35 cases had a rash, of which 9 cases (25.71%) had painful rashes, predominantly in the genital or anal region, accounting for 65.71% (23/35). Eleven cases (31.43%) of monkeypox cases were co-infected with Human Immunodeficiency Virus (HIV), with a 48.57% (17/35) prevalence of sexually transmitted diseases. Throat swabs and vesicle swabs were collected from all cases for nucleic acid testing, with a positive detection rate of 62.86% (22/35)in throat swabs, median CT value of 31, and 100% (35/35) in vesicle swabs, median CT value of 22. Conclusion The main clinical feature of monkeypox is rash, and the early recognition of skin rash by clinical doctors is key to early detection of cases.The high positive rate of monkeypox nucleic acid detection in vesicle specimens indicates the need for timely nucleic acid testing in high-risk individuals presenting with rash symptoms for early diagnosis and treatment.