DNA Damage in Critically Ill COVID-19 Patients

Covid19 Clinical Trial

Official title:

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

Clinical Trial Details — Status: Completed

Administrative data

• NCT number: NCT04772703
• Other study ID #: CVV_COVID-19_DNA
• Status: Completed
• Start date: November 23, 2020
• Est. completion date: February 28, 2021

Study information

• Verified date: September 2022
• Source: University Hospital Hradec Kralove
• Contact: n/a
• Is FDA regulated: No
• Study type: Observational

Clinical Trial 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.

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.

Recruitment information / eligibility

• Status: Completed
• Enrollment: 10
• Est. completion date: February 28, 2021
• Est. primary completion date: February 28, 2021
• Accepts healthy volunteers: No
• Gender: All
• Age group: 18 Years and older

Eligibility

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

Related Conditions & MeSH terms

• COVID-19
• Covid19
• DNA Damage
• Multiple Organ Failure
• Organ Failure, Multiple

Intervention

Diagnostic Test:
• COMET ASSAY
The description in the detailed description of the study

Locations

Country	Name	City	State
Czechia	University Hospital Hradec Kralove	Hradec Králové	Trebeš

Sponsors (1)

Lead Sponsor	Collaborator
University Hospital Hradec Kralove

Country where clinical trial is conducted

Czechia, 

Outcome

Type	Measure	Description	Time frame	Safety issue
Primary	single-stranded DNA breaks	Changes in the single-stranded DNA breaks in peripheral lymphocytes	One week
Primary	oxidation of DNA bases	Changes in oxidised purine and pyrimidine bases of peripheral lymphocytes DNA	One week
Secondary	Mortality	All cause 30-day mortality	One month