Mozambique has a robust HIV control program and currently conducts HIV vaccine trials. One measure of a vaccine's success is development of antibodies, the vaccine-induced seropositivity(VISP). This study assesses the understanding and practices regarding VISP from trial participants and healthcare providers, as well as ethical challenges encountered during the informed consent. We conducted in-depth, semi-structured interviews using open and closed-ended questions with three categories of participants: healthcare professionals [n = 26], researchers [n = 5] and trial participants [n = 20]. For qualitative data, content analysis was performed and thematic axes were assigned. For quantitative analysis, descriptive analysis was performed. Trial participants had a better understanding of VISP's implications than front-line healthcare professionals. 53.8% healthcare professionals interviewed, stated that they would test the trial participants using an HIV rapid test, and initiate antiretroviral therapy immediately if positive. VISP may result in HIV misdiagnoses for trial participants when its broader pratical implications are not well understood.