Characterization of Enterococci

Completed

• Conditions: Samples From Patients With Enterococcal Infections

• Registration Number: NCT04777240
• Lead Sponsor: Sohag University

Brief Summary

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

Detailed Description

Not available

Study Timeline

• Study Start: 2020-02-01
• Status Verified: 2021-02
• Study First Submitted: 2021-02-20
• Study First Submitted QC: 2021-02-25
• Primary Completion: 2021-02-28
• Study First Posted: 2021-03-02
• Study Completion: 2021-03-10
• Last Update Submitted: 2021-04-05
• Last Update Posted: 2021-04-06

Recruitment & Eligibility

• Status: COMPLETED
• Sex: All
• Target Recruitment: 52

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 & Design

• Study Type: OBSERVATIONAL
• Study Design: Not specified

Primary Outcome Measures

Name	Time	Method
IV)Phenotypic detection of virulence marker of Enterococci (hemolytic activity)	1 week	by Detection of hemolytic activity on the blood agar
V) Phenotypic detection of virulence marker of Enterococci(Caseinase production)	2 weeks	by Detection of Caseinase production on Muller hinton agar containing skimmed milk 3%.
VIII) Antibiotic sensitivity test:	3 monthe	1. 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.; 2. 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.
IX) Molecular Identification of commonest Enterococcus species	1 month	by conventional gene specific uniplex PCR for E. faecalis and E. Faecium
X) Molecular detection of virulence genes	2 months	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.
VII) Phenotypic detection of virulence marker of EnterococcI (Biofilm formation)	2 weeks	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
II) Identification of Enterococci:	2 month	by inoculation on Bile esculin azide \& growing colonies further identified by : 1. microscopically after staining by gram stain; 2. catalase test; 3. grow on high concentration of NaCl (6.5%)
I) Sample collection	2 months	Samples are collected from intensive care unit (ICU) including urine, pus swabs, sputum, blood, tracheal aspirates and pharyngeal swabs
III) Phenotypic detection of virulence marker of Enterococci(Gelatinase activity)	2 weeks	by culture on nutrient agar containing gelatin.Positive results appeared as liquefaction of gelatin
VI) Phenotypic detection of virulence marker of Enterococci(Formation of Slime layer)	1 week	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.

Trial Locations

Locations (1)

Sohag Faculty of Medicine; 🇪🇬 Sohag, Egypt