1. Sample: urine, pus and sputum samples.
2. Culture: the samples will be inoculated on the MacConkey agar and subcultured on eosine methylene blue agar.
3. Identification of isolates will be done by:
* Colony morphology.
* Gram staining.
* Biochemical reactions: including Indole, Methyl red, Voges-Proskauer, and Citrate utilization (IMViC) tests.
4. Antimicrobial susceptibility testing of the isolates will be performed using the Kirby-Bauer method (disc diffusion method) using Muller Hinton agar according to the Clinical and Laboratory Standards Institute (CLSI 2025) guidelines for selected groups of antibiotics.
5. Phenotypic detection of biofilm formation by the microtitre plate method.
6. Phenotypic detection of efflux pump activity by ethidium bromide-agar cartwheel method.
7. Detection of biofilm genes (fimH-1, mrkA, and mrkD) by conventional PCR.
8. Detection of efflux pump genes (acrAB, tolC, and mdtk) by conventional PCR.