Rabies is a viral zoonotic disease of public health importance in India. The disease is usually transmitted by a rabid animal bite, scratches and licks on broken skin and mucous membrane. Rabies is reported from all continents except Antarctica although prevalence is more in Asia and Africa. India is endemic for rabies virus (RABV) throughout the country except for islands of Lakshadweep, Andaman & Nicobar. In the present study, brain samples of clinically suspected animals obtained during the RABV outbreak at National Zoological Park, New Delhi, India, in 2016 were initially tested for rabies antigen by DFAT. All the 32 DFAT positive brain samples were then processed for RNA isolation. The extracted RNA was amplified for glycoprotein (G) gene of 590 bp by RT-PCR followed by nucleotide sequencing. Nucleotide alignment revealed only substitutions without any insertion or deletion. Amino acid alignment revealed replacement at four positions namely L244I, T264A, M310I and N375K. The amino acid change at position where Leucine is replaced by Isoleucine has not been observed in available RABV references of the G gene region either from India or other countries. This mutation was observed in all the samples included in the present study. A phylogenetic tree showed that all the study sequences belonged to Arctic like 1a lineage, in RABV genotype 1. In conclusion, RABV in the study region is acquiring genetic variation; which may have effect on the pathogenicity. This indicates that continued molecular surveillance of the circulating rabies virus strains should be carried out
