A total of 438 urine specimens were collected from Rizgary Teaching hospital in Erbil, Iraq, from September to December 2017. Extended spectrum beta lactamase was determined using the Vitek-2 automated system and confirmed by using the combination disk diffusion test. Biofilm formation was checked using 96-well flat bottom microtiter plates. Results: Out of the 438 urine specimens, only 37.89% (n =166) developed an infection, the most common isolate was Escherichia coli. The distribution of the bacterial species according to the patients' gender found to be significant (P = 0.014). The proportion of betalactamase producing isolates was 29% (n = 31). The strength of biofilm formation among Klebsiella species was significantly higher than in Escherichia coli (P <0.001), while a non-significant difference (P = 0.163) was observed between beta lactamase production and ability of biofilm formation. The sensitivity and specificity of VITEK-2 in the detection of extended-spectrum beta-lactamase were 79.48% and 80.95%, respectively. Conclusion: The study revealed that Klebsiella species were stronger biofilm producers. Beta lactamase producing isolates do not have a greater ability of biofilm formation.
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