Molecular Detection Of Efflux Pump and Virulence Factors Genes in Pseudomonas Aeruginosa

NCT05642767 | Unknown DMC

• 1 other identifier: Soh-Med-22-11-18
• Study Type: observational
• Target: 75
• Locations: 1 country
• Sites: 1

Brief Summary

Pseudomonas aeruginosa (PA) is a ubiquitous aerobic, non-fermentative Gram-negative rod that is widely associated with nosocomial pneumonia and can lead to severe illness with poor outcomes, particularly in critically ill people due to the ability of some strains to cause lung epithelial injury and spread into the circulation. 2 In the intensive care unit, PA infection is ranked among the top five causes of the bloodstream, pulmonary, surgical site, urinary tract, and soft tissue infections.

Conditions

Pseudomonas Aeruginosa

Keywords

Efflux genes, Pseudomonas , virulence, PCR

Outcome Measures

Primary Outcomes (3)

• Isolation and identification of pseudomonas aeruginosa using culture and automated system techniques

  identification of pseudomonas aeruginosa in different clinical samples collected from Sohag University hospital using different laboratory techniques as culture on citramide agar, Staining with Gram, biochemical reactions such as Oxidase test, sugar fermentation test, and automated identification using vitek2 system

  1 December 2022 to 1 February 2023

• Identification of recent antibiotic sensitivity pattern using Modified Kerby -Disc Diffusion method

  Determination of recent antibiotic sensitivity pattern using different antibiotics by disc diffusion method by spreading the inoculum in pitry dish containing Muller Hinton Agar, then different discs containing antibiotics are placed at a distance of 1.5 cm, then incubated at 37 co for 24 hrs. The diameter of the zone of inhibition is measured to determine MIC for each antibiotic according to the guidelines of CSLI 2022.

  1 December 2022 to 1 February 2023

• Molecular Identification of some virulence factors and efflux genes using PCR

  Molecular detection of some virulence factors and efflux genes using specific primers by conventional PCR. primers of the following genes will be used as exoS,exoU, toxA, mex A, mex B. Extraction of DNA will be done first, followed by amplification technique using the thermal cycler. Detection of amplified DNA will be done using Agrose gel electrophoresis stained with ethidium bromide.

  1 February 2023 to 30 March 2023

Study Arms (2)

Patients with pseudomonas aeruginosa infections

All patients suffer from infections that can be caused by pseudomonas aeruginosa. Clinical Data will be obtained as: * Data about clinical manifestations including fever, expectoration, pus from wounds, urinary symptoms, symptoms of upper respiratory tract infections, and symptoms of otitis externa. * Samples will be cultured on cetrimide agar. * Antibiotic sensitivity testing will be done by disc diffusion method according to CLSI. * Molecular detection to efflux genes and some virulence genes by PCR.

Diagnostic Test: culture on cetrimide agar; Diagnostic Test: Staining with Gram stain; Diagnostic Test: Antibiotic sensitivity testing; Diagnostic Test: Molecular detection to efflux genes and some virulence genes

Patients with infections other than pseudomonas aeruginosa

* Data about clinical manifestations including fever, expectoration, pus from wounds, urinary symptoms, symptoms of upper respiratory tract infections, and symptoms of otitis externa. * Samples will be cultured on different culture media and automated identification by Vitek system.

Diagnostic Test: culture on cetrimide agar; Diagnostic Test: Staining with Gram stain; Diagnostic Test: Antibiotic sensitivity testing

Interventions

culture on cetrimide agar DIAGNOSTIC_TEST

Samples will be inoculated on cetrimide agar using the plating out technique.

Patients with infections other than pseudomonas aeruginosa; Patients with pseudomonas aeruginosa infections

Staining with Gram stain DIAGNOSTIC_TEST

colonies on cetrimide agar will be spread on glass slide and stained by gram stain

Patients with infections other than pseudomonas aeruginosa; Patients with pseudomonas aeruginosa infections

Antibiotic sensitivity testing DIAGNOSTIC_TEST

Antibiotic sensitivity testing will be done by disc diffusion method according to CLSI

Patients with infections other than pseudomonas aeruginosa; Patients with pseudomonas aeruginosa infections

Molecular detection to efflux genes and some virulence genes DIAGNOSTIC_TEST

Molecular detection to efflux genes and some virulence genes by conventional PCR

Patients with pseudomonas aeruginosa infections

Eligibility Criteria

• Age: 4 Weeks - 80 Years
• Sex: all
• Healthy Volunteers: No
• Age Groups: Child (0-17), Adult (18-64), Older Adult (65+)
• Sampling Method: Non-Probability Sample

Study Population

This study will be carried out on all patients suffering from infections that can be caused by pseudomonas aeruginosa. Samples (pus, urine, blood, sputum, ear discharge) will be collected from different departments at sohag university hospital. Clinical Data will be obtained as: \- Data about clinical manifestations including fever, expectoration, pus from wounds, urinary symptoms, symptoms of upper respiratory tract infections, and symptoms of otitis externa.

You may qualify if:

• All patients suffering from infections that can be caused by pseudomonas aeruginosa

You may not qualify if:

• Samples diagnosed to have organisms other than pseudomonas aeruginosa.

Sponsors & Collaborators

• Sohag University: lead

Study Sites (1)

Sohag University

Sohag, Egypt

RECRUITING

Related Links

• Related Info

Biospecimen

Retention: SAMPLES WITH DNA

Samples (pus, urine, blood, sputum, ear discharge) will be collected from different departments at sohag university hospital.

MeSH Terms

Conditions

Pseudomonas Infections; Pathologic Complete Response

Interventions

Staining and Labeling

Condition Hierarchy (Ancestors)

Gram-Negative Bacterial Infections; Bacterial Infections; Bacterial Infections and Mycoses; Infections; Disease Progression; Disease Attributes; Pathologic Processes; Pathological Conditions, Signs and Symptoms

Intervention Hierarchy (Ancestors)

Histocytological Preparation Techniques; Cytological Techniques; Clinical Laboratory Techniques; Diagnostic Techniques and Procedures; Diagnosis; Histological Techniques; Investigative Techniques

Study Officials

• Mohamed H Alrawy

  Faculty of medicine, Sohag university

  STUDY CHAIR

• Ebtisam M Gad

  Faculty Of Medicine, Sohag university

  STUDY CHAIR

Central Study Contacts

Noha S Shafik, lecturer

CONTACT

01067261504; Nohasaber@med.sohag.edu.eg

Nesma A Mohammed, lecturer

CONTACT

01006780725; Nesmaaateef@med.sohag.edu.eg

Study Design

• Study Type: observational
• Observational Model: CASE CONTROL
• Time Perspective: CROSS SECTIONAL
• Sponsor Type: OTHER
• Responsible Party: PRINCIPAL INVESTIGATOR
• PI Title: lecturer of Medical Microbiology and Immunology, faculty of medicine

Study Record Dates

• First Submitted: November 19, 2022
• First Posted: December 8, 2022
• Study Start: December 1, 2022
• Primary Completion: March 30, 2023
• Study Completion: May 1, 2023
• Last Updated: April 12, 2023

Record last verified: 2023-04

Data Sharing

• IPD Sharing: Will not share

Locations

EG (1)