Female sex workers (FSW) represent a focal point of the HIV epidemic in India. HIV self-testing (HIVST) could mitigate under-diagnosis of HIV and reduce disease transmission in this population. This study assessed the acceptability of HIVST through focus group discussions (FGD) with FSW. FSW expressed willingness to use HIVST and preference for saliva-based HIVST over blood-based HIVST and preferred that HIVST education, administration, and storage take place in trusted community centers and not in brothels. We provide preliminary recommendations for the implementation of an acceptable and feasible HIVST program for FSW in India.