Background Mobile health (mHealth) can be beneficial in monitoring the complex healthcare regimen for people with HIV that includes adhering to medication and refraining from risky practices such as unsafe sex and injection drug use. Not only is mHealth often implemented without appropriate feasibility and acceptability research, but there is limited mHealth research among key HIV-positive populations such as men who have sex with men (MSM) and transgender women (TGW). Methods This study assessed access to and use of mobile technology and acceptability of mHealth among 150 HIV-positive MSM and TGW who were prescribed antiretroviral therapy (ART) in Malaysia-an emerging economy with rapid telecommunications growth and societal stigma against these groups. Results Findings among the 114 MSM and 36 TGW reveal high levels of depression (42%), stigma (2.53/4.00) and risky sexual behavior (30%), and suboptimal ART adherence (22%). On the other hand, the sample had excellent access to smartphones (75.3%) and the internet (78%), and had high acceptance of mHealth especially for those with suboptimal ART adherence. Conclusion In settings like Malaysia where homosexuality and cross-dressing are socially and legally stigmatized, HIV prevention and treatment strategies delivered using an mHealth platform have the potential to overcome in-person barriers.