Malaria is the most widespread endemic disease in Cameroon, and asymptomaticPlasmodium(gametocyte) carriers (APCs) constitute more than 95% of infectious human reservoirs in malaria endemic settings. This study assesses the knowledge of asymptomatic malaria (ASM) among health care workers (HCWs) in health facilities (HFs) in the Centre Region of Cameroon and the acceptability of usingArtemisia afratea to treat APCs. A cross-sectional descriptive survey was conducted among 100 HCWs, in four randomly selected HFs in the Centre Region, in the period of 1–20 April 2022, using semi-structured self-administered questionnaires. Logistic regression analyses were performed to determine factors associated with knowledge. More than seven in eight (88%) respondents were aware of the existence of ASM, 83% defined ASM correctly, 75% knew how it was diagnosed, 70% prescribe ACTs for APCs, and 51.1% were informed about ASM transmission. The professional category of HCWs was significantly associated with their knowledge of the existence and transmission of ASM, and longevity of service was associated with knowledge of transmission (p< 0.05). Two-thirds (67%) of respondents knew aboutArtemisia afratea, 53.7% believed that it was effective in treating ASM, and 79% were willing to prescribe it if authorized. There was a fair level of knowledge of ASM among HCWs in the study settings.