BackgroundIt is estimated that sub-optimal feeding, especially non-exclusive breastfeeding in the first 6months of life, results in 1.4million deaths and 10% of disease burden in children younger than five years. Worldwide, it is estimated that only 34.8% of infants are exclusively breastfed for the first 6months of life, the majority receiving some other food or fluid in the early months. Besides, the Ethiopian demographic and health survey (2016) stated that the median duration of exclusive breastfeeding in Tigray region was 3.8 months which is shorter than the recommended duration. The main purpose of this study was to determine the magnitude of exclusive breastfeeding practice and associated factors among HIV positive mothers in public hospitals of Tigray region, Northern Ethiopia.MethodsA facility based cross-sectional study was conducted from July 9 to October 11, 2016, in public hospitals of Tigray region. Data was collected by using structured questionnaire using face-to-face interview among 304 eligible women through a systematic sampling technique. Data was analyzed using SPSS version 20.0. Binary and multiple variable logistic regressions (“odds ratio”) analyses were calculated with 95% CI and p value ≤ 0.05 as significance were used.ResultTwo hundred seventy (88.8%) of mothers practiced exclusive breastfeeding for the first six months of life. Infant feeding counseling during antenatal care of last pregnancy [AOR = 6.9, 95% CI; 2.63, 17.99], knowledge on exclusive breastfeeding (AOR = 5.5, 95% CI; (2.12, 14.02] and attitude towards exclusive breastfeeding [AOR = 7.9; 95% CI; 2.96, 21.21] had significant association with exclusive breastfeeding practice.ConclusionsA high proportion of mothers practiced exclusive breastfeeding for the first six months of life. Infant feeding counseling, knowledge and attitude towards exclusive breastfeeding practice were the predictors of exclusive breastfeeding among HIV positive mothers. Strengthening infant feeding counseling during antenatal care and improving mothers’ knowledge and attitude on exclusive breastfeeding is essential.