Antibodies Responses to COVID-19 Infection in Hospitalized Patients

NCT04520880 | Unknown DMC

• 1 other identifier: ICU-06072020
• Study Type: observational
• Target: 158
• Locations: 1 country
• Sites: 1

Brief Summary

1.5. Why this clinical study? The prevalence of seropositivity following SARS-CoV 2 infection might have its own potential benefits in terms of predicting the end of pandemic and the validity of herd immunity. It is not clear if SARS-CoV 2 infection would have a long-lasting antibody-mediated immunity, and if the antibodies' persistence is dependent on disease severity.depends on the severity of illness. If evidence is provided about the persistence of antibodies that is reflective of the protective immune response, serodiagnosis will be an important tool to identify individuals with various risk for infection, and those who are in need of receiving the forthcoming vaccines. The here proposed prospective clinical study will test the prevalence of seropositivity following SARS-CoV 2 infection in critically ill patients compared to those who do not require intensive care unit (ICU) admission or invasive ventilation with respect to the IgM and IgG levels.

Conditions

Hospitalized Patients; Laboratory-confirmed SARS-CoV 2 Infection; Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV 2 Infection)

Outcome Measures

Primary Outcomes (2)

• Changes in the levels of S specific antibodies in severely ill patients compared to mild cases.

  The measurements are dependent on epitope recognitions for synthetic, adsorbed S proteins

  Changes from baseline (4 to 6 weeks) at 16 weeks after the onset of symptoms of SARS-Cov2 infection

• Changes in the levels of N specific antibodies in severely ill patients compared to mild cases.

  The measurements are dependent on epitope recognitions for synthetic, adsorbed N proteins

  Changes from baseline (4 to 6 weeks) at 16 weeks after the onset of symptoms of SARS-Cov2 infection

Secondary Outcomes (10)

• S specific binding antibodies of SARS-CoV-2

  4 to 6 weeks and 16 weeks after the onset of symptoms of SARS-Cov2 infection

• N specific binding antibodies of SARS-CoV-2

  4 to 6 weeks and 16 weeks after the onset of symptoms of SARS-Cov2 infection

• Neutralizing antibodies directed against S protein of SARS-CoV-2

  4 to 6 weeks and 16 weeks after the onset of symptoms of SARS-Cov2 infection

• The severity category of critically ill

  Day 0, 4 to 6 weeks and 16 weeks after the onset of symptoms of SARS-Cov2 infection

• Length of ICU

  For 16 weeks after the onset of symptoms of SARS-Cov2 infection

Study Arms (2)

Critically ill SARS-Cov2 patients

Critically ill patients with acute hypoxemic respiratory failure defined as those admitted to ICU and receiving mechanical ventilation (invasive or non-invasive) or high-level supplemental oxygen (via a high-flow nasal cannula or non-rebreathing face mask at a flow rate of 15 L per min or greater), at or during hospitalization

Diagnostic Test: Testing procedure for Binding antibodies; Diagnostic Test: Neutralizing antibodies

Hospitalized non-critically ill SARS-Cov2 symptomatic patients

Hospitalized non-critically ill SARS-Cov2 symptomatic patients

Diagnostic Test: Testing procedure for Binding antibodies; Diagnostic Test: Neutralizing antibodies

Interventions

Testing procedure for Binding antibodies DIAGNOSTIC_TEST

Levels of S-specific and N-specific binding antibodies will be measured by enzyme immune-sorbent assay (ELISA). Briefly, a purified, recombinant S or N proteins will be coated into 96-well plates and incubated for 1 hour at room temperature. The plates will be then washed with 1X PBS five times. The unbound regions of the coated S proteins are blocked by 5% non-fat dry milk. After overnight incubation at 4 C. The plates are washed several times with 5X PBS. Patients sera samples are added in duplicate and incubated for 1 hour at room temperature. After several washes. anti-human IgG HRP conjugate is added. After 1-hour incubation at room temperature, the plates are washed with 1X PBS five times 3,3', 5,5' tetramethylbenzidine (TMB) substrate is added to each wells and incubated for 5 minutes. The reaction will be then stopped with 0.2 M sulfuric acid and the absorbance is measured at 450nm.

Critically ill SARS-Cov2 patients; Hospitalized non-critically ill SARS-Cov2 symptomatic patients

Neutralizing antibodies DIAGNOSTIC_TEST

Neutralizing antibodies against the S protein of SARS-CoV-2 are measured by PV microneutralization assay. Briefly, individual plasmids expressing S.fl gene, luciferase genes, and reporter genes are co-transfected into 293T cells to produce luciferase-expressing pseudo-viruses. Then, pseudo-viruses are added at equal volume into 96 well plates and incubated at 37 for 1 hour. Then, the ACE2 expressing 293T cells are added into the 96well pates with 100 ul of patients' sera in duplicates. After, a single round of replication the cells are lysed with lysis buffer. Luciferase activity is measured by the luciferase assay kit. The activity of luciferases is proportional to the number of cells infected. Luciferase activity is then measured by the luminometer device and the neutralization rate is calculated.

Critically ill SARS-Cov2 patients; Hospitalized non-critically ill SARS-Cov2 symptomatic patients

Eligibility Criteria

• Age: 18 Years+
• Sex: all
• Age Groups: Adult (18-64), Older Adult (65+)
• Sampling Method: Non-Probability Sample

Study Population

Symptomatic inpatients (ICU and none ICU) patients with laboratory-confirmed SARS-CoV 2 infection admitted to the participating centers will be screened, eligible patients will be consented then assessed for the timing of the onset of symptoms. Patients will be included for testing of the virus-specific antibodies levels if they spend at least 2 weeks from symptoms onset.

You may qualify if:

• Symptomatic infection with at least 2 weeks' time interval since the symptom's onset.
• Laboratory-confirmed COVID-19 with positive SARS-CoV-2 real-time polymerase chain reaction (RT-PCR) testing of nasopharyngeal or oropharyngeal swabs.
• Inpatients in either isolated wards or ICU.
• Agreement for blood sampling during the course of the study.

You may not qualify if:

• Decline consent to participate.
• Pregnancy.
• Asymptomatic patients, who are PCR positive during routine screening upon admission
• Administration of immunoglobulins within the 3 proceeding months including Covid-19 convalescent plasma.
• Patients with Don't resuscitate orders
• Patients with terminal illnesses regardless of the severity of SARS-CoV 2 infection.

Sponsors & Collaborators

• Imam Abdulrahman Bin Faisal University: lead
• Dammam Medical Complex: collaborator
• Institute for Research and medical consultations (IRMC): collaborator

Study Sites (1)

Imam Abdulrahman Bin Faisal University

Dammam, Eastern Province, 31952, Saudi Arabia

Location

Biospecimen

Retention: SAMPLES WITHOUT DNA

3-5 ml of blood would be obtained from each patient will be collected at 4 to 6 weeks and 16 weeks intervals after the onset of symptoms of SARS-Cov2 infection in a tube. The blood samples are centrifuged and sera will be separated and stored at 20 C or below until further immunological testing. Levels of S-specific and N-specific binding antibodies will be measured by enzyme immune-sorbent assay (ELISA). Briefly, a purified, recombinant S or N proteins will be coated into 96-well plates and incubated for 1 hour at room temperature.

MeSH Terms

Conditions

COVID-19

Condition Hierarchy (Ancestors)

Pneumonia, Viral; Pneumonia; Respiratory Tract Infections; Infections; Virus Diseases; Coronavirus Infections; Coronaviridae Infections; Nidovirales Infections; RNA Virus Infections; Lung Diseases; Respiratory Tract Diseases

Study Officials

• Mohammed S Alshahrani, MD

  Critical Care and emergency Department, Associate Professor, College of Medicine, Imam Abdulrahman Ben Faisal University

  PRINCIPAL INVESTIGATOR

• Iman Almansour, PhD

  Associate Professor, Department of epidemic disease research, IRMC, IAU

  PRINCIPAL INVESTIGATOR

• Mohamed R El Tahan, MD

  Anesthesiology Department, Associate Professor, College of Medicine

  STUDY DIRECTOR

Central Study Contacts

Mohammed S Alshahrani, MD

CONTACT

+966 55 696 6663; msshahrani@iau.edu.sa

Mohamed R El Tahan, MD

CONTACT

+966569371849; mohamedrefaateltahan@yahoo.com

Study Design

• Study Type: observational
• Observational Model: OTHER
• Time Perspective: PROSPECTIVE
• Sponsor Type: OTHER
• Responsible Party: SPONSOR

Study Record Dates

• First Submitted: July 21, 2020
• First Posted: August 20, 2020
• Study Start: August 1, 2020
• Primary Completion: December 1, 2020
• Study Completion: February 1, 2021
• Last Updated: August 20, 2020

Record last verified: 2020-08

Data Sharing

• IPD Sharing: Will share
• Shared Documents: STUDY PROTOCOL, SAP, ICF, CSR
• Time Frame: There is an indefinite time limit, following publishing the scientific paper
• Access Criteria: The database will be locked as soon as all data are entered, and all discrepant or missing data are resolved - or if all efforts are employed and we consider that the remaining issues cannot be fixed. At this step, the data will be reviewed before database locking. After that, the study database will be locked and exported for statistical analysis. At this stage, permission for access to the database will be removed for all investigators, and the database will be archived.

Locations

SA (1)