Biomedical waste materials produced in hospitals and clinics usually harbour infectious microorganisms, which can pose a threat to societal health and well-being, especially when improperly disposed of. Escherichia coli O157:H7 is a very virulent pathogen of man and animals, with a high prevalence due to a very low infective dose. The present study was conducted between July to September, 2021 on biomedical waste in General Hospital Dutse to detect Escherichia coli O157:H7. A total of 200 samples were randomly collected from the different units of the hospital and analysed using standard microbiological, biochemical and molecular techniques. The frequency of occurrence of Escherichia coli was 73 (36.5%) out of which Escherichia coli O157:H7 were 20 (10%) and others were 127 (63.5%). The detected Escherichia coli O157:H7 was subjected to tests for the virulence genes stx1 and stx2, Four (4) of the samples were found to possess the virulent gene stx2 alone while the other had neither. The presence of presumptive Escherichia coli O157:H7 in the waste samples may directly point to improper waste management practices and may result into outbreak of disease is the waste materials are not disposed of sanitarily. It is therefore recommended that more attention should be placed on the waste management practices employed in the hospital and a critical review of the already existing practices to tally with international standards.