Infection transmission in radiology departments is a very important concern for radiology staff. This cross-sectional study aimed to assess the knowledge, attitude, and practice (KAP) about infection prevention and control among diagnostic radiographers in Yemen. An online questionnaire was conducted between February and March of 2022. A total of 378 diagnostic radiographers participated in this study and completed the questionnaire. The results revealed that more than half of the participants had adequate knowledge (51.3%) and appropriate practice (51.6%), whereas more than two-thirds of participants (73%) had positive attitude toward infection prevention and control. The findings indicate a significant difference in the KAP scores in infection prevention with respect to the age, educational status, and occupational experience of the radiographers, while there was no significant difference between the KAP scores of radiographers due to the gender. Furthermore, the radiographers’ KAP scores were positively correlated. This study showed that the diagnostic radiographers in Yemen have inadequate levels of knowledge and practice regarding infection prevention and control. Therefore, the implementation of infection-control educational programs is recommended to improve their understanding and practice for a better infection control effect.