This work reports a simplistic approach to develop an antimicrobial coating material based on natural rubber latex (NRL). Here, crosslinking of the NRL (cis 1,4-poly(isoprene) was carried out utilizing “thiol-ene chemistry” using trimethylolpropane tris(3-mercaptopropionate) (trithiol) as a crosslinker and Irgacure-184 as a photoinitiator under UV irradiation. This process was utilized to prepare an antimicrobial NRL coated cotton fibre via adopting a dip coating method and subsequent curing. Prior to the coating on the cotton fibre and curing of NRL, it was mixed with modified chitosan (Cts) stabilized silver nanoparticles (Ag NPs) at a pH 7.4 (physiological pH). Presence of Ag NPs incorporated cured NRL coating over the cotton fibre, significantly increases its elongation at break up to 25% as compared to the pure cotton fibre (14%). It also increases the hydrophobicity (Water contact angle = 102°) of the fibre. The as-prepared fibre exhibits superior antimicrobial activity against both Gram-negative and Gram-positive bacteria and non-cytotoxic tested over the NIH 3T3 fibroblast cell line which indicates its potential application in the antimicrobial textile.