Magister Curationis - MCur / Early infant diagnosis (EID) programmes offer diagnosis of HIV, which facilitates provision of life-saving care to infants infected with HIV. Implementing programmes for EID and treatment has proved challenging in Malawi. Many infants access EID late or not at all. Previous studies have shown that lack of knowledge among health care providers (HCPs) is a challenge to effective EID. Little is known on the knowledge and skills of health care providers in Malawi. Aim: The aim of the study was to investigate the knowledge and skills of HCPs on EID of HIV in Mzuzu, Malawi. Objectives: (i) to examine the knowledge of HCPs on EID of HIV; and (ii) to determine the skills of HCPs on EID of HIV. Methods: A descriptive cross-sectional survey design with a quantitative approach was used. The study was conducted in three hospitals in Mzuzu, Malawi. The population was HCPs (doctors, nurses/midwives, clinical officers and medical assistants) working in maternity, paediatric wards and under-five clinics. A total of 68 HCPs participated in the study. A closed-ended self-administered questionnaire was used to collect data. Data were analysed using the Statistical Package for Social Science version 23. Descriptive statistics were used to present the frequency tables of observations. Ethical approval was sought from the University of the Western Cape Senate Research Committee and Malawi National Health Research Council. Results: The results on the knowledge of HCPs demonstrate that 38% of them had a score of <69% (poor), 25% scored within 70–79% (fair), and 37% scored >80% (good). Results on the skills showed that 69% of the HCPs scored <69% (poor), 15% scored within 70–79% (fair), and 16% scored >80% (good). The results also showed a correlation between the knowledge of HCPs and their level of education achievement (certificate, diploma and degree) as well as the skills of HCPs and their department of work. Conclusion: The study found that more than one-third of the HCPs lacked knowledge and skills on EID of HIV. These findings reflect the need to address the practical challenges of EID service delivery. Recommendations: There is a need to increase the efforts that are being put in place to train HCPs on EID of HIV in order to scale up EID. Training should assess the needs of HCPs regarding the knowledge and skills required in the delivery of EID services.