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Objectives To examine the feasibility of non-occupational postexposure prophylaxis (nPEP) as a national strategy for HIV prevention in China, we investigated nPEP usage and related sociodemographic and behavioural factors among five key populations at high risk of contracting HIV. Methods We conducted a cross-sectional study among five key populations from November 2018 to September 2019 in China using convenience sampling to recruit participants aged >= 18 years, self-reporting HIV status as either negative or unknown and providing written informed consent. Univariable and multivariable logistic regression models were fitted. Results Our analysis included data from 2022 participants with a mean age of 35 years (SD=11.62). Only 57 (2.82%) participants had ever used nPEP. Sociodemographic and behavioural factors related to nPEP usage included populations (p<0.0001), age (p<0.05), education (p<0.05), nPEP knowledge (p<0.01), receiving conventional HIV prevention services (p<0.05) and HIV testing (p<0.05). A significant percentage (26%) of nPEP users used nPEP medication more than once. Challenges and concerns, such as multiple use of nPEP and syndemic conditions, were emerging. Conclusions Key populations in China had low nPEP usage rates. Female sex workers, people who use drugs, older and illiterate individuals with poor nPEP knowledge, not using HIV prevention services or never tested for HIV should be emphasised. Implementing nPEP services would be an important way to access high-risk individuals for intensive and tailored HIV prevention and intervention. Challenges of providing nPEP services and future study foci are highlighted.