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Background: Continued assessment of nurses' knowledge and attitude is necessary for improving their performance at workplace. We attempted to evaluate nurses' knowledge and attitude toward HIV-infected patients and their underlying factors., Methods: We conducted a descriptive, cross-sectional study participated by 218 nurses and collected relevant data., Results: Majority (57.3%) of the nurses had 1 to 5 years of experience in the nursing job, and 78.4% of them were involved in the provision of direct care to HIV-infected individuals for ≤5 years, and only 6.4% for ≥11 years. The overall knowledge and attitude of the participants were good (3.25/5 and 3.42/5 scores, respectively). The knowledge regarding appropriate care of HIV-infected patients was significantly better among the registered nurses compared to nursing assistants (NA) with limited years of formal education (odds ratio [OR] = 0.37; 95% confidence interval [CI] = 0.20-0.69; P <.001). In the provision of care to the HIV-infected patients, the attitude of female participants was significantly better than the males (unadjusted OR = 0.18; 95% CI = 0.05-0.60; P = .01)., Conclusion: Inclusion of relevant HIV- and AIDS-related topics in the curriculum for NAs and closely guided hands-on training of the nurses are likely to improve nurses' knowledge and attitude toward the provision of care to HIV-infected individuals.