Multidrug Resistant Klebsiella Pneumoniae Infection
Conditions
Brief summary
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.
Detailed description
K. pneumoniae is a member of the Gram negative Enterobacteriaceae family. It is a rod-shaped, capsulated, non-motile, non-spore-forming bacteria characterized by a unique mucoid look on agar plates. Klebsiella pneumoniae is an opportunistic pathogen that is responsible for a wide range of community-acquired and healthcare-associated infections including urinary tract infections, wound infection, bacteremia, and pneumonia 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.
Interventions
GENETICGenotypic Detection of Biofilm Formation and Efflux Pump Activity.
1. Phenotypic detection of biofilm formation by the microtitre plate method. 2. Phenotypic detection of efflux pump activity by ethidium bromide-agar cartwheel method. 3. Detection of biofilm genes (fimH-1, mrkA, and mrkD) by conventional PCR. 4. Detection of efflux pump genes (acrAB, tolC, and mdtk) by conventional PCR.
Sponsors
Sohag University
Study design
Observational model
CASE_ONLY
Time perspective
CROSS_SECTIONAL
Eligibility
Sex/Gender
ALL
Healthy volunteers
Yes
Inclusion criteria
1-Age and sex range: any age and any sex. 2-Clinical setting: hospital-acquired infections. 3-Single pathogen focus. 4-MDR K. pneumoniae which develop resistance to at least one agent from three or more antibiotic classes. \-
Exclusion criteria
1-Recent antibiotics : Use within 48-72 hours before enrollment. 2-Mixed infections. \-
Design outcomes
Primary
| Measure | Time frame |
|---|---|
| Prevelace of biofilm formation genes in MDR Klebsiella pneumonea isolates (%) | 6 months |
Secondary
| Measure | Time frame |
|---|---|
| Prevelace of efflux pump genes in MDR Klebsiella pneumonea isolates (%) | 6 months |
Contacts
CONTACTGhofran Mahrous Mohammed
Outcome results
None listed