Characterization of Enterococci

Characterization of Enterococci; Distribution of Virulence Markers, Virulence Genes and Antibiotic Resistance Pattern of the Isolated Species

Study on Characterization of Enterococci because nowadays it become an important cause of nosocomial infections .detection of the most common two species of Enterococci and most common virulence factors & its genes with determination of antibiotics sensitivity test for the isolated strains

Study Overview

• Status: Completed
• Conditions: Samples From Patients With Enterococcal Infections

• Study Type: Observational
• Enrollment (Actual): 52

Contacts and Locations

Study Locations

• Egypt
  • Sohag, Egypt
    • Sohag faculty of medicine

Participation Criteria

Eligibility Criteria

• Ages Eligible for Study: 2 months to 90 years (Child, Adult, Older Adult)
• Accepts Healthy Volunteers: N/A
• Genders Eligible for Study: All

• Sampling Method: Probability Sample

Study Population

patients in ICU in Sohag University Hospital

Description

Inclusion Criteria:

• any patients in ICU has manifestations of urinary tract infections, surgical wound infections, intra-abdominal infections, intrapelvic infections, bacteraemia and infective endocarditis

Exclusion Criteria:

• patients receiving antibiotics in previous 48 hours

Study Plan

How is the study designed?

What is the study measuring?

Primary Outcome Measures

Outcome Measure	Measure Description	Time Frame
I) Sample collection	Samples are collected from intensive care unit (ICU) including urine, pus swabs, sputum, blood, tracheal aspirates and pharyngeal swabs	2 months
II) Identification of Enterococci:	by inoculation on Bile esculin azide & growing colonies further identified by : microscopically after staining by gram stain; catalase test; grow on high concentration of NaCl (6.5%)	2 month
III) Phenotypic detection of virulence marker of Enterococci(Gelatinase activity)	by culture on nutrient agar containing gelatin.Positive results appeared as liquefaction of gelatin	2 weeks
IV)Phenotypic detection of virulence marker of Enterococci (hemolytic activity)	by Detection of hemolytic activity on the blood agar	1 week
V) Phenotypic detection of virulence marker of Enterococci(Caseinase production)	by Detection of Caseinase production on Muller hinton agar containing skimmed milk 3%.	2 weeks
VI) Phenotypic detection of virulence marker of Enterococci(Formation of Slime layer)	Formation of Slime layer by culture on Brain heart infusion agar containing 5% sucrose, plates are incubated for 24 hrs at 37oC. Positive strains gave mucoid and slimy colonies.	1 week
VII) Phenotypic detection of virulence marker of EnterococcI (Biofilm formation)	using microtiter plate reader. Biofilm formation is scored as nonbiofilm forming (-), weak - (+), moderate - (++), and strong - (+++) corresponding to the A630 values ≤1, 1-≤2, 2-≤3, and>3, respectively	2 weeks
VIII) Antibiotic sensitivity test:	by 1- disc diffusion test using Vancomycin 30 μg, Teichoplanin 30 μg, Tetracycline 30 μg, Ampicillin 10 μg and Erythromycin 15 μg. Results ,Results are interpreted according to CLSI 2018.; E-test: MICs (minimal inhibitory concentrations) of Vancomycin are measured by E-test for confirmation of vancomycin resistance among the isolated Enterococci. Results are interpreted according to CLSI guidelines.	3 monthe
IX) Molecular Identification of commonest Enterococcus species	by conventional gene specific uniplex PCR for E. faecalis and E. Faecium	1 month
X) Molecular detection of virulence genes	Identification of virulence genes; gel E (gene for gelatinase), asa1 (gene for aggregation substance), cylA (gene for cytolysin activator), esp (gene for Enterococcal surface protein) Hyl (gene for Hyaluronidase) of E. faecalis and E. faecium was performed by uniplex PCR.The amplified products are visualized on 2% agarose gel stained with ethidium bromide. The stained gels are visualized and documented with a gel documentation system and analyzed visually to determine the size of PCR amplicons of the target genes directly by comparison with 100 bp DNA ladder marker.	2 months

Collaborators and Investigators

• Sponsor: Sohag University
• Investigators: Principal Investigator: Ekram Ab Rahmand Mahmoud, lecturer, Sohag faculty of medicine

Study record dates

Study Major Dates

• Study Start (Actual): February 1, 2020
• Primary Completion (Actual): February 28, 2021
• Study Completion (Actual): March 10, 2021

Study Registration Dates

• First Submitted: February 20, 2021
• First Submitted That Met QC Criteria: February 25, 2021
• First Posted (Actual): March 2, 2021

Study Record Updates

• Last Update Posted (Actual): April 6, 2021
• Last Update Submitted That Met QC Criteria: April 5, 2021
• Last Verified: February 1, 2021

More Information

Terms related to this study

• Other Study ID Numbers: Soh-Med-21-02-19

Plan for Individual participant data (IPD)

• Plan to Share Individual Participant Data (IPD)?: NO

Drug and device information, study documents

• Studies a U.S. FDA-regulated drug product: No
• Studies a U.S. FDA-regulated device product: No