BACKGROUND: Female sex workers (FSWs) and long distance truck drivers (LDTDs) are considered key populations at high risk for HIV transmission due to high prevalence. The intersection of these mobile populations presents unique challenges in the fight against HIV and the movement towards reducing new infections. OBJECTIVE: The purpose of this study was to explore the nature of sex trade along a particular transport route. Sexual transactions and the vulnerabilities that exist between these two groups with regards to HIV/AIDS are described, with the purpose of furthering the agenda for targeted interventions. METHODS: Qualitative in-depth interviews were conducted with 14 participants, seven FSWs and seven LDTDs. We recruited FSWs through snowballing, and LDTDs through intercepts at truck stops. Semi-structured interview guides were used for data collection, and thematic analysis was conducted. RESULTS: The sex trade in this study is characterized by competition, fuelled by money-driven and age-disparate rivalry. Despite widespread HIV knowledge, FSWs contend with persistent challenges regarding condom use negotiation, induced by more money in the exchange for unsafe sex. Despite the placement of wellness centres in truck stops along the highway, LDTDs face stigma related challenges with regards to testing for HIV and personal acknowledgement of their involvement in the sex trade. CONCLUSION: The nature of the sex trade along the highway continues to be risky despite the availability of HIV testing and antiretroviral treatment (ART). The sex trade is perceived to be increasing along trucking routes, in spite of measures instituted to limit access to FSWs. FSWs struggle to cope with the pressure of unprotected sex because of the need to generate more income, as well as avoid incidents of violence and threats. Interventions along transport routes need to be inclusive of FSWs who could play a vital role in stigma reduction amongst LDTDs through peer education.