In Australia, approximately 30% of people diagnosed with HIV are not accessing treatment and 8% of those receiving treatment fail to achieve viral suppression. Barriers limiting effective care warrant further examination. This mixed-methods systematic review accessed health and social sector research databases between November and December 2015 to identify studies that explored the perspective of people living with HIV in Australia. Articles were included for analysis if they described the experiences, knowledge, attitudes and beliefs, in relation to treatment uptake and adherence, published between January 2000 and December 2015. Quality appraisal utilised the Mixed Methods Appraisal Tool Version 2011. Seventy-two studies that met the inclusion criteria were reviewed. The interplay of lack of knowledge, fear, stigma, physical, emotional and social issues were found to negatively impact treatment uptake and adherence. Strategies targeting both the individual and the wider community are needed to address these barriers.