In this cross-sectional study, blood samples were collected from 423 pregnant women and from 55 healthy volunteers who visited the University of Gondar Hospital. Serum concentration of vitamin A was measured by high performance liquid chromatography.After controlling for total serum protein, albumin and demographic variables, the mean ± SD serum vitamin A in HIV seropositive pregnant women (0.96 ± 0.42 μmol/L) was significantly lower than that in pregnant women without HIV infection (1.10 ± 0.45 μmol/L, P < 0.05). Likewise, the level of serum vitamin A in HIV seropositive non-pregnant women (0.74 ± 0.39) was significantly lower than that in HIV negative non-pregnant women (1.18 ± 0.59 μmol/L, P < 0.004). VAD (serum retinol < 0.7 μmol/L) was observed in 18.4% and 17.7% of HIV infected and uninfected pregnant women, respectively. Forty six percent of non-pregnant women with HIV infection had VAD while only 28% controls were deficient for vitamin A (P = 0.002).The present study shows that VAD is a major public health problem among pregnant women in the tropical settings of Northwest Ethiopia. Considering the possible implications of VAD during pregnancy, we recommend multivitamin (which has a lower level of vitamin A) supplementation in the care and management of pregnant women with or without HIV infection.Vitamin A deficiency (VAD) is known to be a significant public health problem around the world and it is particularly serious among women of reproductive age in South-East Asia and Africa [1-4]. It has now become evident that VAD in women has negative consequences on their health status as well as on their infants [3,4]. The link between VAD morbidity and mortality from infectious diseases  and non-infectious diseases [6-8] has been known for several years.VAD in pregnant women is associated with night blindness, severe anaemia, wasting, malnutrition, and reproductive and infectious morbidity , and increased risk of mortality 1-2 years following delivery .