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

Обзор

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.

Подробное описание

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.

Вмешательства

• Генная терапия 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.

Первичные конечные точки

• Prevelace of biofilm formation genes in MDR Klebsiella pneumonea isolates (%) [Срок оценки: 6 months]

Вторичные конечные точки (1)

• Prevelace of efflux pump genes in MDR Klebsiella pneumonea isolates (%) [Срок оценки: 6 months]

Критерии участия

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.

\-

Критерии приведены из реестра в оригинале (на английском). Окончательную оценку соответствия проводит исследовательский центр.

Здоровые добровольцы: Да

Дизайн исследования

Модель наблюдения

Только случаи

Центры проведения

Список центров уточняется — проверьте первичный протокол.

Идентификаторы

NCT: NCT07541690 · Soh-Med--25-12-4MS

Первоисточники (государственные реестры)

• ClinicalTrials.gov (NIH, США)

Открыть это исследование на ClinicalTrials.gov ↗