Objective: To document the causes and clinical significance of direct Coomb’s test distal convoluted tubule in neonatal Jaundice at a tertiary care setup. Study Design: Cross-sectional study. Place and Duration of the Study: Fatima Memorial Hospital, Lahore Pakistan, from Nov 2019 to Oct 2020. Methodology: All babies born to “O” or Rh-negative blood group mothers were included. In addition, data including demographics, mother’s blood group, baby’s blood group, direct Coomb’s test results, bilirubin values, and need for treatment was recorded. Results: The study included 989 neonates. Of all, 489(49.4%) were male, the mean birth weight was 2.80±0.75 kg, and the mean gestation was 36±4.3 weeks. ABO mismatch was present in 452(45.7%) cases and Rh mismatch in 123(12.4%). Of all, 58(5.9%) had positive direct Coomb’s test. Amongst the positive direct Coomb’s test patients, 44(75.8%) babies needed treatment, of which 40(90.9%) received phototherapy, and 4(6.8%) needed Intensive phototherapy (360 degrees). No baby needed an exchange transfusion. Conclusion: In most cases, positive direct Coomb’s test was due to ABO blood group incompatibility. In addition, early phototherapy can control bilirubin rise in most cases, so few would need aggressive or intensive treatment.Keywords: ABO mismatch, Direct Coomb’s test, Neonatal jaundice, Rh mismatch.