In the present study, a low-cost filter material such as Nylon was used for the disinfection of groundwater poses an excellent bacterial filtration activity against Escherichia coli (gram-negative) cultures. Textile fabrics have gained wide acceptance as a filtration membrane, and various substrates having a pore size in microns such as Nylon, polyester, and cotton have been shown to develop microbial filtration properties. In this study, a simple method to filter bacteria using nylon is presented and the solutions are prepared using Autoclave and Laminar Air Flow Chamber. For checking the bacterial concentration in the underground water, a very small volume of sample was tested by keeping it in shaking Incubator. The results revealed that nylon filters were able to decrease the concentration of E. coli from a groundwater sample, with a higher removal efficiency achieved by using two membranes and a lower efficiency by using a single membrane. This study, therefore, suggests that the Nylon membrane demonstrated antibacterial activity against E.Coli. For the disinfection of groundwater, Nylon membrane can be used as a potential alternative cost-effective filter for production of safe drinking water.