Voluntary counselling and testing for Human Immunodeficiency Virus (HIV) and Acquired Immune Deficiency Syndrome (AIDS) has always been one of the key policy interventions in the management and control of HIV/AIDS transmission. However, the prevalence of HIV testing among reproductive women in the Gambia remains low despite near universal information about HIV and Sexually Transmitted Infections (STIs) in the Gambia. Understanding factors influencing HIV testing uptake provides empirical data for the development of targeted evidenced-based strategies aimed at enhancing HIV testing uptake. Therefore, this study examined the factors associated with HIV testing among reproductive women aged 15–49 years in the Gambia. Data on weighted sample of 11,865 women from the 2019–2020 Gambia Demographic and Health Survey were analyzed in this study. Chi square, bivariate and multivariate logistic regression models were fitted and analysis conducted through Complex Samples Analysis in Statistical Package for Social Sciences (SPSS). Level of significance was set at p < 0.05 and 95% CI. Further analysis was conducted to determine the variability in HIV testing among women stratified by rural and urban centers. Prevalence of HIV testing among reproductive women was 42.1% (95% CI = 40.1–44.2%) in the Gambia. Women aged 20–24 years and 25–29 years (aOR = 3.10, 95% CI = 2.51–3.83) and (aOR = 4.52, 95% CI = 3.61–5.54) were more likely to test for HIV than those aged 15–19 years, respectively. Married women (aOR = 5.90, 95% CI = 4.84–7.02) were more likely to test for HIV compared to those who were not in any union. Respondents with higher education in urban centers (aOR = 2.65, 95% CI = 2.08–3.86) were likely to test for HIV compared to those in rural areas. HIV testing in the Gambia among reproductive women is low. Age, marital status, wealth index, place of residence, educational level, recent sexual activity, previous history of risky sexual behaviors, and history of an STI were associated with HIV testing. Health interventions targeted at increasing HIV testing uptake should factor in these.