Background: The organic acids like succinic acid and malic acid have an antibacterial impact on E.coli infection. Objective: The aim of the present study is the detection of E.coli in urine samples using PCR and antibiotic susceptibility testing of strains obtained from people and food using ampicillin and erythromycin, in addition to testing malic and succinic acids against uropathogenic E.coli. Material and methods: This study describes a biochemical and polymerase chain reaction E.coli test. PCR can accurately verify E.coli colonies on culture plates. The Minimum Inhibitory Concentration (MIC) method assessed organic acids and antibiotics for bactericidal action. Results: The study showed that malic acid has a higher ratio of inhibition that increase from 0.06 to hit 0.14 in concentration 0.3 mg/ml, while Ampicillin''s ratio of inhibition fluctuates the same level until rise gradually to reach 0.04 in concentration 0.09 mg/ml. Succinic acid''s ratio increased to 0.35 in concentration 0.4 mg/ml, while in 0.5 mg/ml hit 0.4 and dropped to 0.1 in the concentration of 0.6 mg/ml, in contrast to  Erythromycin that have a higher ratio of inhibition increased gradually  to reach 1.5 in concentration 0.7 mg/ml. Conclusion: Urinary tract infections caused by E.coli may be challenging to treat empirically in Ile-Ife due to antibiotic resistance. In order to reduce the selection pressure that could lead to the spread of E. coli in the environment and to increase the likelihood of a successful treatment, culture and susceptibility tests must be performed prior to administering any antibiotics.