DNA Damage in Critically Ill COVID-19 Patients

Sponsor

University Hospital Hradec Kralove (Other)

Overall Status

Recruiting

CT.gov ID

NCT04772703

Collaborator

(none)

Enrollment

Location

8.2

Anticipated Duration (Months)

1.2

Patients Per Site Per Month

Study Details

Study Description

Brief Summary

Critically ill patients with COVID-19 are exposed to high oxidative stress which is potential harm to the DNA. Peripheral lymphocytes' DNA will be investigated using the comet assay on changes in oxidative damage to the purine and pyrimidine bases and single-stranded DNA breaks.

Condition or Disease	Intervention/Treatment	Phase
Covid19 Organ Failure, Multiple DNA Damage	Diagnostic Test: COMET ASSAY

Detailed Description

Data from hospital documentation will be recorded for all patients: demographic data, clinical data (Horowitz index, the necessity of pronation position, opening maneuvers, connection to ECMO, state of the circulation and other organ functions), laboratory data: parameters of inflammatory activity and COVID-19 specific markers (blood count with differential leukocyte count, CRP, IL-6, ferritin, procalcitonin, D-dimers).

In the study, 4 ml of heparinized peripheral blood will be collected for analytical purposes beyond standard practice for lymphocyte isolation and monitoring of oxidative DNA damage upon admission to the ICU (D1), after 2 days (D3) and on day 7 (D7) of the hospitalization. The same blood sample will be used to determine EPA and DHA omega-3 in erythrocyte membranes. At the same intervals and frequency, 10 ml of urine will be collected for the determination of 8-hydroxy-2-deoxyquanosine (8-OHdG), so a total of 30 ml of urine will be collected for analytical purposes.

Details for analytical processing: peripheral blood lymphocyte damage will be assessed using the alkaline COMET ASSAY method for DNA breakage and modified COMET ASSAY using specific enzymes for the detection of oxidized bases, endonuclease III (ENDO III) for the detection of oxidized pyrimidines and formamidopyrimidine DNA glycosylase (FPG) for the detection of oxidized purines. Leukocyte repair capacity will be determined using a modified COMET ASSAY, which uses extracts of patient lymphocytes and HeLa cells with defined DNA damage. The extent of 8-hydroxy-2-deoxyquanosine (8-OHdG) DNA damage in patients' urine will be determined by determining the concentration of 8-OHdG by ELISA.

Study Design

Study Type:

Observational

Anticipated Enrollment :

10 participants

Observational Model:

Cohort

Time Perspective:

Prospective

Official Title:

DNA Damage in Critically Ill Patients With SARS-CoV-2 Infection With Organ Failure

Actual Study Start Date :

Nov 23, 2020

Anticipated Primary Completion Date :

Feb 28, 2021

Anticipated Study Completion Date :

Jul 30, 2021

Outcome Measures

Primary Outcome Measures

single-stranded DNA breaks [One week]
Changes in the single-stranded DNA breaks in peripheral lymphocytes
oxidation of DNA bases [One week]
Changes in oxidised purine and pyrimidine bases of peripheral lymphocytes DNA

Secondary Outcome Measures

Mortality [One month]
All cause 30-day mortality

Eligibility Criteria

Criteria

Ages Eligible for Study:

18 Years and Older

Sexes Eligible for Study:

All

Accepts Healthy Volunteers:

Inclusion Criteria:

critically ill adult patient admitted to ICU with positive RT-PCR test or bed-side antigen test on COVID-19 with signs and symptoms of organ failure
informed consent

Exclusion Criteria:

rejection by the patient to sign an informed consent

Contacts and Locations

Locations

	Site	City	State	Country	Postal Code
1	University Hospital Hradec Kralove	Hradec Králové	Třebeš	Czechia	50005

Sponsors and Collaborators

University Hospital Hradec Kralove

Investigators

None specified.

Study Documents (Full-Text)

None provided.

More Information

Publications

None provided.

Responsible Party:

University Hospital Hradec Kralove

ClinicalTrials.gov Identifier:

NCT04772703

Other Study ID Numbers:

CVV_COVID-19_DNA

First Posted:

Feb 26, 2021

Last Update Posted:

Feb 26, 2021

Last Verified:

Feb 1, 2021

Individual Participant Data (IPD) Sharing Statement:

Plan to Share IPD:

Studies a U.S. FDA-regulated Drug Product:

Studies a U.S. FDA-regulated Device Product:

Additional relevant MeSH terms:

COVID-19

Multiple Organ Failure

Study Results

No Results Posted as of Feb 26, 2021