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Background: Human immunodeficiency virus (HIV) infection constitutes a major medical complication of pregnancy and is associated with adverse feto-maternal outcomes. However, the relationship between maternal serum selenium levels and pregnancy outcomes has been inconsistent. Objective: This study aimed to determine the relationship between maternal serum selenium status and pregnancy outcome in HIV-positive and HIV-negative women in a tertiary health facility. Methodology: A comparative cross-sectional study was carried out among HIV-positive and HIV-negative pregnant women at a tertiary health-care facility in Owerri. Participants were recruited from the labor ward and interviewed using a structured questionnaire. One hundred and ten HIV-positive pregnant women were compared with an equal number of HIV-negative pregnant women. They were matched for age, parity and gestational age. Selenium level was measured using atomic absorption spectrophotometer. Maternal packed cell volume (PCV) was also assessed at recruitment. At delivery, the birth weight was measured using a standard weighing scale and documented. Cases of preterm births, perinatal deaths, major congenital abnormalities, and neonatal admission were noted and also documented. Statistical analysis was performed using means and standard deviation. Chi-square test, Student's t-test, logistic regression, and Pearson correlation were also employed. Statistical significance was considered at P < 0.05. Results: HIV-positive pregnant women had significantly lower mean serum selenium concentration compared with HIV-negative pregnant women (64.3 ± 19.6 μg/L vs. 100.1 ± 30.9 μg/L; P < 0.001). There was a statistically significant association between serum selenium concentration and birth weight among both HIV-positive and HIV-negative pregnant women ( P < 0.001). Similarly, a statistically significant association was seen between serum selenium and maternal PCV in HIV-positive and HIV-negative pregnant women ( P = 0.024 and P < 0.001, respectively). However, there was no association found between serum selenium and other pregnancy outcomes. Conclusion: HIV-positive pregnant women had a lower mean serum selenium level compared to HIV-negative pregnant women. There was a significant association between low maternal serum selenium level and maternal anemia, as well as low birth weight, especially in HIV-positive pregnant women.