BackgroundMultidrug-resistant (MDR)E. coliwith extended-spectrum β-lactamases (ESBLs) is becoming endemic in health care settings around the world. Baseline data on virulence and AMR of specific lineages ofE. colicirculating in developing countries like India is currently lacking.MethodsWhole-genome sequencing was performed for 60 MDRE. coliisolates. Genome-wide analysis was performed at single nucleotide polymorphism (SNP) level resolution to identify the relation between the isolates in context of time, virulence and AMR determinants possessed.ResultsGenome comparison revealed the presence of ST-131 global MDR and ST410 as emerging-MDR clades ofE. coli. AMR gene profile for cephalosporin and carbapenem resistance differed between the clades. GenotypesblaCTX-M-15andblaNDM-5were common among cephalosporinases and carbapenemases, respectively. For aminoglycoside resistance,rmtBwas positive for 31.7% of the isolates, of which 30% were co-harboring carbapenemases. Further, the FimH types and virulence gene profile positively correlated with the SNP based phylogeny, which also revealed the evolution of MDR clones among the study population with temporal accumulation of SNPs. The predominant clone was ST167 (blaNDMlineage) followed by ST405 (global clone ST131 equivalent) and ST410 (fast spreading high risk clone).ConclusionsThis is the first report on the whole genome analysis of MDRE. colilineages circulating in India. Data from this study will provide public health agencies a baseline portfolio of AMR and virulence in pathogenicE. coliin the region.