Bacterial infections caused by antibiotic resistance has resulted in bacteremia, low body immunity and caused millions of deaths in different parts of the country. The spread of Carbapenem and colistin resistance in Gram-negative bacteria has emerged as a global health concern across India. The aim of this work was to determine the prevalence of carbapenem and colistin resistance among the clinical isolates of Tamil Nadu. For this work, clinical samples were collected from a diagnostic center in Chennai, Tamil Nadu. A total of 226 isolates were collected belonging to 11 genera. All isolates were identified at species level by 16S rRNA sequencing. The isolates include Acinetobacter baumanni (n=12), Citrobacter spp (n=8), E.coli (n=8), Enterobacter cloacae (n=8), Klebsiella Oxytoca (n=8), Morganella morganii (n=8), Klebsiella pneumoniae (n=8), Pseudomonas aeruginosa (n=79), Proteus mirabilis (n=8), Proteus vulgaris (n=8), Salmonella typhi (n=8), Strenotophomonas maltophia (n=8), Sphingomonas pauchmobilis (n=8). Out of the 226 isolates, 124 (54%) and 3 (1.3%) were found to be resistant to Meropenem and Colistin. The most prevalent beta-lactamase gene was blaOXA-48 (n=4), followed by blaNDM (n=3). One strain of Pseudomonas aeruginosa (PA-9) carried both blaOXA-48 and blaNDM. Plasmid incompatibility (inc/rep) typing results showed that the resistance genes were present in the isolates carrying plasmid types IncA/C, IncFIA-FIB  and IncFIIA. The plasmid borne resistance genes in Pseudomonas aeruginosa that carried the blaOXA-48 like showed transfer of plasmid to E.Coli AB1157. This work highlights the importance of Carbapenem resistance and acquisition of Carbapenamase genes on plasmids in the Gram-negative bacterial isolates.