BackgroundHIV is a major public health problem, especially in developing countries including Ethiopia. Exploring the spatial pattern, distribution and associated factors of HIV Seropositivity is important to monitor, and design effective intervention programs. Therefore, this study showed associated factors and spatial variation of HIV Seropositivity in Ethiopia.MethodThis secondary data analysis, sampling technique and procedures were done by Ethiopipan Central Statistical Agency. A total sample of 25,774 individuals data were extracted from the 2016 EDHS data mainly HIV biomarkers, IR, MR, and GPS. Spatial heterogeneity analysis were done using tools like Morans I, Local G*, Interpolation, and Kulldorff’s scan statistic. The spatial analysis was carried out by using open source software (QGIS, GeoDa, SaTScan). Multilevel logistic regression analysis was used to identify both individual and household level factors associated with HIV Seropositivity and the analysis was carried out by Stata 14. Finally AOR with 95% confidence interval of mixed-effect logistic regression result in the full model was used to report.ResultThe prevalence of HIV/AIDS was found 0.93% at the national level. The highest prevalence regions were Gambela, Addis Ababa, Harari, and Dire Dawa, which accounts for 4.79%, 3.36%, 2.65%, and 2.6%, respectively. Similarly, the most likely high-risk HIV Seropositivity spatial cluster was found in the Gambela region and Addis Ababa followed by Harari and Diredawa. In a multilevel analysis at the individual level being married [AOR = 2.19 95%CI: (1.11, 4.31)] and previously married [AOR = 6.45, 95%CI (3.06, 13.59)] were significantly associated with serropositivity. Regarding household level place of residence [urban: AOR = 6.13 CI: (3.12, 12.06)] were associated with HIV Seropositivity.ConclusionThe distribution of HIV cases was not random. High cluster HIV cases were found in Gambela, Addis Abeba, Harari, and Dire Dawa. At the individual level, some characters are high risk like being previously married, start sex at a younger age, female household headed, urban residence, and lower household size is more affected by HIV/AIDS. So any concerned bodywork around this risk group and area can be effective in the reduction of transmission.