Multi-drug-resistant (MDR) strains of biofilm-producing Escherichia coli are being reported worldwide and are threatening the health of human beings. These species are seen as the highest priority for the development of new phage biocontrol agents. Therefore, we isolated and characterized new and effective lytic phages as biocontrol agents against MDR E. coli. Aim:-The aim of this study is in vitro lytic efficacy of phages against MDR Escherichia coli Methodology: This study was conducted in department of Microbiology at RKDF Medical College Hospital & Research Centre, SRK University, Bhopal, Madhya Pradesh.This study included 140 Escherichia coli .Out of 140 Escherichia coli 122 were MDR Escherichia coli and rest were normal. Bacteriophages (phages) were isolated from different wastewater samples and subjected for plaque formation. Result: The result of this study revealed that best plaque formation was seen at 1012pfu/ml concentration. Conclusion:- In conclusion, it may be suggested bacteriophage therapy might be beneficial for the patient who are suffering with the MDR strains from a long period of time