Fear of acquiring HIV infection among healthcare workers can impede the quality of prevention of mother-to-child transmission (PMTCT) services. Objectives This paper focuses on understanding healthcare workers’ fear of acquiring HIV infection and aims to identify the key contributing factors associated with that fear while providing PMTCT related services to pregnant women in Fiji. Additionally, this paper highlights the implications of this fear on providing quality PMTCT related service to pregnant women. Design Qualitative methodology was employed in this study. Setting Three tertiary hospitals and associated health centers in Fiji. Study Sample A total of 58 healthcare providers were interviewed, including doctors (n=12), midwives (n=19), nurses (n=14), laboratory technicians (n=5) and counsellors (n=8) were interviewed, using in-depth interviews. Methods We collected the data through in-depth interviews conducted in Fiji between April-May 2013 and February-March 2014. Thematic analysis was used to analyse the data. Results We found that fear of occupationally acquired HIV infection was prevalent among healthcare workers, especially among nurses and midwives. Needlestick injuries, lack of awareness of infection control process, shortage of personal protective equipment, and delayed presentation of pregnant women with unknown HIV status in labour ward were identified as the main contributory factors associated with the fear. Conclusion Our findings would be valuable in developing strategies to overcome the fear among healthcare workers. Simple intervention such as standardised infection prevention and control training would be valuable for mitigating the fear of HIV infection among healthcare workers.