Background: The array of β–lactamases like ESBLs and AmpC produced by Gram negative bacilli causes many somber infections like neonatal sepsis and their increasing incidence major health concern. Aim: To determine the incidence of ESBLs and AmpC β–lactamase producing Escherichia coli and Klebsiella spp isolated from neonatal sepsis cases. Methodology: Blood samples from 382 clinically suspected neonatal septicaemic cases were processed by standard microbiological methods. Antimicrobial susceptibility testing was carried out by Kirby Bauer disk diffusion method as per CLSI guidelines. Screening and confirmation of ESBLs production was also done according to CLSI guidelines (2015). Disc antagonism test (DAT) and E test (Ezy MICTM strip) tests were used for detection of AmpC. Results: Of the 382 blood samples, 124 (32.46%) samples showed bacterial growth. Culture positivity in males was 34.36% and 29.67% in females. Culture positivity among the EONS and LONS cases was found to be 47.38% and 52.42%. There was preponderance of Gram negative isolates (58.87%) over Gram positive isolates (37.9%) and Candida albicans (3.23%). E. coli (41.09%) was mostly isolated from EONS cases while Klebsiella spp (31.51%) was isolated almost equally from both types of neonatal sepsis. The incidence of E. coli and Klebsiella spp was found to be 43.33% and 47.83% respectively. Pure AmpC as well as co–production of ESBL and AmpC was seen in 6.25% E. coli and 7.69% among Klebsiella spp. Conclusion: It is very important to differentiate between ESBLs and AmpC producers so that the treatment guidelines can be wisely formulated. E strip test as well as disk antagonism test was found to be useful in detecting AmpC production.