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Phenotypic and Genotypic Detection of Biofilm Formation and Efflux Pump Activity in Multi-drug Resistant Klebsiella Pneumoniae

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Summary

NIH's ClinicalTrials.gov registered observational study NCT07541690, an examination of phenotypic and genotypic mechanisms of antibiotic resistance in multi-drug resistant Klebsiella Pneumoniae. The study will analyze 70 urine, pus, and sputum samples using culture, biochemical testing, antimicrobial susceptibility profiling, and PCR-based gene detection for biofilm formation and efflux pump activity. No compliance obligations are created by this registry entry.

“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.”

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What changed

This document is a clinical trial registry entry for study NCT07541690, which will examine phenotypic and genotypic mechanisms underlying multi-drug resistance in Klebsiella Pneumoniae. The study protocol involves culture-based identification, antimicrobial susceptibility testing per CLSI 2025 guidelines, phenotypic detection of biofilm and efflux pump activity, and conventional PCR detection of specific resistance genes. Registry entries on ClinicalTrials.gov do not create regulatory obligations for healthcare providers, laboratories, or pharmaceutical companies. Researchers studying similar antimicrobial resistance mechanisms may find this protocol relevant to their own study design.

Affected parties include clinical microbiology researchers, infectious disease specialists, and antimicrobial resistance surveillance programs. The study does not impose reporting requirements, testing mandates, or compliance deadlines on external entities. No penalties or enforcement mechanisms are associated with this registry entry.

Archived snapshot

Apr 22, 2026

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Phenotypic and Genotypic Detection of Biofilm Formation and Efflux Pump Activity in Multi-drug Resistant Klebsiella Pneumoniae

Observational NCT07541690 Kind: OBSERVATIONAL Apr 21, 2026

Abstract

  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.
    1. 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.
    2. Phenotypic detection of biofilm formation by the microtitre plate method.
    3. Phenotypic detection of efflux pump activity by ethidium bromide-agar cartwheel method.
    4. Detection of biofilm genes (fimH-1, mrkA, and mrkD) by conventional PCR.
    5. Detection of efflux pump genes (acrAB, tolC, and mdtk) by conventional PCR.

Conditions: Multidrug Resistant Klebsiella Pneumoniae Infection

Interventions: Genotypic Detection of Biofilm Formation and Efflux Pump Activity.

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Last updated

Classification

Agency
NIH
Published
April 21st, 2026
Instrument
Notice
Branch
Executive
Legal weight
Non-binding
Stage
Final
Change scope
Minor

Who this affects

Applies to
Clinical investigators Healthcare providers
Industry sector
5417 Scientific Research
Activity scope
Clinical research Antimicrobial susceptibility testing Microbiology study design
Geographic scope
United States US

Taxonomy

Primary area
Public Health
Operational domain
Clinical Operations
Topics
Pharmaceuticals Healthcare

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