Phenotypic and Genotypic Detection of Biofilm Formation and Efflux Pump Activity in Multi-drug Resistant Klebsiella Pneumoniae
For patients and families
In plain language
An automatic summary of structured registry data. It is an orientation aid, not a substitute for the official protocol or a physician assessment.
- What is being studied
- The protocol lists: Genotypic Detection of Biofilm Formation and Efflux Pump Activity..
- Who it may be relevant to
- Registry conditions: Multidrug Resistant Klebsiella Pneumoniae Infection. Basic parameters: No limits · All.
- What needs checking
- Age, condition and sex are only basic indicators. Prior treatment, laboratory values and other mandatory requirements appear in the eligibility criteria below.
- Where it takes place
- Center list to be confirmed — check the primary protocol.
- Next step
- Save the trial, show it to the treating physician, and confirm current recruitment with the study center. Costs, documents and travel →
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Overview
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
- Genetic Genotypic 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.
Primary outcome measures
- Prevelace of biofilm formation genes in MDR Klebsiella pneumonea isolates (%) [Time frame: 6 months]
Secondary outcome measures (1)
- Prevelace of efflux pump genes in MDR Klebsiella pneumonea isolates (%) [Time frame: 6 months]
Eligibility criteria
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.
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Exclusion Criteria:1-Recent antibiotics : Use within 48-72 hours before enrollment.
2-Mixed infections.
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Criteria are shown verbatim from the registry (in English). Final eligibility is always assessed by the study center.
Healthy volunteers: Yes
Study design
- Observational model
- Case-only
Study locations
Center list to be confirmed — check the primary protocol.
Identifiers
NCT: NCT07541690 · Soh-Med--25-12-4MS