Kothi-identified men who have sex with men in India are highly marginalized and are at high-risk for HIV. This study examines HIV testing among 132 self-reported HIV-negative and unknown serostatus kothis recruited from public sex environments in Chennai, India. Using logistic regression we identified variables associated with HIV testing uptake (i.e., being tested and knowing the result). Sixty-one percent reported HIV testing uptake. At the bivariate level, married men, those with low HIV transmission knowledge, those who engaged in unprotected anal sex and unprotected receptive anal sex were at lower odds of reporting testing uptake. In multivariate analysis, married men and those with low levels of HIV transmission knowledge were at decreased odds of being tested, as were kothis who experienced forced sex. Culturally competent programs engaging married kothis are needed. Interventions to facilitate HIV prevention education and systemic interventions to combat sexual violence may facilitate HIV testing uptake among kothis.