Deficiency of Glucose-6-phosphate dehydrogenase (G6PD) is the most common metabolic disorder of red blood cells leading to hemolytic anemia and neonatal hyperbilirubinemia (NNH). With almost 24 million children born annually in India, it is estimated that atleast 390,000 children suffering from this disorder are born in the country every year. This study was performed to screen the newborn for G6PD deficiency and its incidence and relationship with neonatal hyperbilirubinemia. This study was carried out in the Department of Pathology, Silchar Medical College & Hospital, Silchar in 998 newborn of both sexes from June 2013 to May 2014. Out of 998 newborn, 16 (1.6%) cases were found to have deficiency of G6PD enzyme which included 15 (2.8%) males and 1 (0.2%) female. All 16 (11.5%) cases with G6PD deficiency presented with NNH. Screening for G6PD deficiency should be done in every newborn with jaundice as early diagnosis can reduce the number of complication and prevents future acute haemolytic attacks.