Background <p>The use of human milk is one of the most cost-effective interventions in preventing preterm mortality. Donor human milk (DHM) is the preferred option for feeding infants when mother’s own milk is unavailable. We hypothesized that there is a difference in knowledge, attitude and intention to use DHM between mothers of preterm and term infants in Jos.</p> Methods <p>This was a comparative cross-sectional study of mothers of hospitalized preterm and term neonates in four major hospitals in Jos. Data was collected using an interviewer-administered questionnaire and analysed using IBM SPSS version 25.</p> Results <p>There were a total of 157 mothers (78 preterm and 79 term). Thirty-two (41.0%) mothers of preterm and 22(27.8%) mothers of term infants had good DHM knowledge (<i>x</i>² = 3.020, <i>p</i> = 0.082). Also, 57(73.1%) preterm and 62(78.5%) term mothers had favourable attitude towards DHM (<i>x</i>² = 0.646, <i>p</i> = 0.724). Twenty-nine (37.2%) preterm and 26(32.9%) term mothers intended to use DHM (<i>x</i>² = 0.314, <i>p</i> = 0.575) while 65(83.3%) and 56(70.9%) intended to donate milk (<i>x</i>² = 3.441, <i>p</i> = 0.064) respectively. Good knowledge (aOR = 2.974, 95%CI = 1.348–6.567), favourable attitude towards DHM (aOR = 3.446, 95%CI = 1.208–9.825), intention to donate milk (aOR = 9.004, 95%CI = 1.935–41.895), were significantly associated with intention to use DHM while being Catholic (aOR = 0.227, 95%CI = 0.114–0.974) or Muslim (aOR = 0333, 95%CI = 0.114–0.974) were less likely have intention to use DHM.</p> Conclusion <p>Most mothers in Jos did not have good DHM knowledge and did not intend to use DHM. Mothers of preterms did not differ from their term counterparts in knowledge, attitude and intention to use DHM. Educational programmes are recommended to improve DHM use.</p>