Africa is the most severely affected area, accounting for more than two-thirds of the people living with HIV. In sub-Saharan Africa, more than 85% of new HIV-infected adolescents and 63% of all new HIV infections are accounted for by women. Ethiopia has achieved a 50% incidence rate reduction. However, mortality rate reduction is slow, as the estimated prevalence in 2021 is 0.8%. In sub-Saharan Africa, heterosexual transmission accounts for the majority of HIV infections, and women account for 58% of people living with HIV. Most of these transmissions took place during marriage. Thus, this study aimed to explore the spatial variation of premarital HIV testing across regions of Ethiopia and identify associated factors. A cross-sectional study design was employed. A total of 10223 weighted samples were taken from individual datasets of the 2016 Ethiopian Demographic and Health Survey. STATA version 14 and ArcGIS version 10.8 software's were used for analysis. A multilevel mixed-effect generalized linear model was fitted, and an adjusted prevalence Ratio with a 95% CI and p-value < 0.05 was used to declare significantly associated factors. Multilevel models were compared using information criteria and log-likelihood. Descriptive and spatial regression analyses (geographical weighted regression and ordinary least squares analysis) were conducted. Models were compared using AICc and adjusted R-squared. The local coefficients of spatial explanatory variables were mapped. In spatial regression analysis, secondary and above education level, richer and above wealth quintile, household media exposure, big problem of distance to health facility, having high risky sexual behaviour and knowing the place of HIV testing were significant explanatory variables for spatial variation of premarital HIV testing among married women. While in the multilevel analysis, age, education level, religion, household media exposure, wealth index, khat chewing, previous history of HIV testing,age at first sex, HIV related knowledge, HIV related stigma, distance to health facility, and community level media exposure were associated with premarital HIV testing among married women. Premarital HIV testing had a significant spatial variation across regions of Ethiopia. A statistically significant clustering of premarital HIV testing was observed at Addis Ababa, Dire Dawa, North Tigray and some parts of Afar and Amhara regions. Therefore area based prevention and interventional strategies are required at cold spot areas to halt the role of heterosexual transmission in HIV burden. Moreover, the considering the spatial explanatory variables effect in implementations of these strategies rather than random provision of service would make regional health care delivery systems more cost-effective.