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Background: Klebsiella pneumoniae is a major causative agent of diarrhea and acute enteritis in children under 5 years of age, which plays a major role in the development of harmful carcinogenic biofilms due to its strong colonization capacity. The various cell surface molecules and their various structures were the reason for the development and formation of biofilms of klebsiella pneumoniae . Many topics referred to as biofilms, are associated with many health problems. The ability of K.pnemoniae on the formation of the biofilm, as well as the role of a number of virulence factor genes on its formation and contributes to the stability and survival of K. pnemoniae infections. The biofilm is one of the virulence factors of K. pneumoniae. The membrane represents the bacterial cells aggregating and adhering to the living and non-living surfaces producing a thick layer of the extracellular biofilm that works to protect the cells from phagocytosis by the immune system of the host, eliminating the effect of the epithelial cells, and antibiotics. Objectives: to study the genes responsible for the formation of biofilm by klebsiella pneumoniae. Material and Method: the samples have collected from Children under 5years old from Women’s Hospital for Childbirth , Al Noor Hospital for Children, and Al-Sadiq hospital in Hilla city during the period from September month 2020 to March 2021. The population of study was consist of 1hundred, 30 healthy individual apparently no disease is shown, while the other group are sick and include (80 patients had acute gastroenteritis, 48 of the females, 32 of the males. Results : according to the diagnosis of klebsiella pneumoniae isolates that is based on standard laboratory protocols and confirmation of the diagnosis with the VITEC system, our results showed 31.25% is a Klebsiella was the dominant type in the samples and this activity of Klebsiella was achieved to form biofilms by tube method. Conclusion: The results showed that the Klebsiella bacteria are the strongest in the biofilm formation this based on molecular genes in bacteria such as wbbm, wzm and Mrkd gene.