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Alpha hemolysin is a toxin secreted by Escherichia coli causes cell death by binding with the outer membrane, with subsequent oligomerization of the toxin monomer and water-filled channels. These are responsible for osmotic phenomena, cell depolarization, and loss of vital molecules, leading to its demise. A total of 20 clinical isolates of E. coli were screened for the presence of hlyA gene by PCR. We have observed 60% positivity among our isolates. There are several virulent factors that are associated with urinary tract infections, production of alpha hemolysin one amongst those factors also play a crucial role in pathogenesis of UTI.