Clinical, Virological, Serological and Immunological Characteristics During and Following COVID-19 Hospitalization (COVISERA)

Clinical, Virological, Serological and Immunological Characteristics During and Following COVID-19 Hospitalization: a Prospective Cohort Study

Assessment of the association between the severity of COVID-19 and SARS-CoV-2 NAb titers levels for up to six months following primary infection using a live virus NAb assay. Description of SARS-CoV-2 viral shedding and infectiousness during the first 30 days after infection in a group of unvaccinated hospitalized patients.

Study Overview

• Status: Active, not recruiting
• Conditions: COVID-19; COVID-19 Pneumonia

Detailed Description

The studys primary aim is to assess clinical factors, such as disease severity, associated with neutralizing antibody (NAb) production. Furthermore, the study aims to assess the length of SARS-CoV-2 infectiousness and clinical factors associated with viral load.

Patients 18 years or older hospitalized at Copenhagen University Hospital at North Zealand, Copenhagen, Denmark, May 24th,2020 - May 5th, 2021, were routinely screened for COVID-19 by diagnostic oropharyngeal or tracheal RT-PCR samples taken during admission. Patients with a positive SARS-CoV-2 PCR within 48 hours from hospital admission were offered inclusion, if COVID-19 pneumonia was confirmed.

The following were retrieved from patients' electronic records: comorbidities (Charlson Comorbidity Index),vital signs (Early Warning Score), immunocompromised status, time from symptom onset to admission, oxygen treatment, pharmacological treatment, admission length, death and bacterial co-infection.

Paired oropharyngeal swabs and serum samples were collected at inclusion (day 0), days 3, 7, 10, 14, 17, 24, and 30. Serum samples were, if possible, also collected after three and six months. Follow-up time was six months.

RT-qPCR analysis targeted the SARS-CoV-2 RNA-dependent-RNA-polymerase (RdRp)-helicase gene region and two samples with known viral load were included in each PCR-run for quantification of patient samples. Virus was cultured in African green monkey cells (VERO-E6) with incubation for 3 - 4 days and daily microscopic inspection for cytopathogenic effect (CPE). A total of three passages were made before the the virus was interpreted as non-replicant. Cells with CPE were confirmed by RT-qPCR.

The presence of specific antibodies (Ab) against SARS-CoV-2 in serum was assessed using Wantai total-Ab ELISA according to the manufacturer's instructions (Wantai, Beijing, China).

For the in-house live virus NAb analysis, a 50% cut-off value was calculated from quadruplicate virus and cell control wells included on each plate using the following equation: (average optical density (OD) of virus control wells + average OD of cell control wells)/2. The 50% neutralization titer was calculated as the interpolation of the cutoff value with a four-parameter logistic regression curve fitted for each serum serial dilution. To minimize inter-assay variation, the titers were normalized according to a positive control included on each assay plate.

A linear mxied-effects model was used to assess the association between repeated NAb measurements and clinical variables such as age, gender and disease severity. A linear mixed-effects model was also used to assess the association between repeated viral load measurements and clinical variables.

• Study Type: Observational
• Enrollment (Actual): 47

Contacts and Locations

Study Locations

• Denmark
  • Copenhagen, Denmark, 25482
    • Copenhagen University Hospital at North Zealand

Participation Criteria

Eligibility Criteria

• Ages Eligible for Study: 18 years and older (Adult, Older Adult)
• Accepts Healthy Volunteers: No
• Genders Eligible for Study: All

• Sampling Method: Non-Probability Sample

Study Population

Adult unvaccinated hospitalized patients

Description

Inclusion Criteria:

• positive SARS-CoV-2 specimen from upper or lower respiratory tracts (virological criteria)
• consolidations on the chest X-ray described by a radiologist, treating physician or a physician from the study group (radiological criteria)
• the presence of one or more of the following: fever (temperature ≥38.0°C), new-onset cough, pleuritic chest pain, dyspnea or altered breath sounds on auscultation (clinical criteria)

Exclusion Criteria:

• cognitive impairment prohibiting giving informed consent to participation
• by December 14th, 2020, and onwards patients if the time since symptom onset was > seven days at the time of inclusion

Study Plan

How is the study designed?

Design Details

• Observational Models: Cohort
• Time Perspectives: Prospective

What is the study measuring?

Primary Outcome Measures

Outcome Measure	Measure Description	Time Frame
Neutralizing Antibody Titer - Day 0	Baseline Log10 Spike protein neutralizing antibody titer	Day 0
Change from baseline in Neutralizing Antibody Titer - Day 30	Log10 Spike protein neutralizing antibody titer	Day 30
Change from baseline in Neutralizing Antibody Titer - Day 90	Log10 Spike protein neutralizing antibody titer	Day 90
Change from baseline in Neutralizing Antibody Titer- Day 180	Log10 Spike protein neutralizing antibody titer	Day 180
Viral culturing - Day 0	Number of successful viral culturing attempts (SARS-CoV-2)	Day 0
Viral culturing - Day 3	Number of successful viral culturing attempts (SARS-CoV-2)	Day 3
Viral culturing - Day 7	Number of successful viral culturing attempts (SARS-CoV-2)	Day 7
Viral culturing - Day 10	Number of successful viral culturing attempts (SARS-CoV-2)	Day 10
Viral culturing - Day 14	Number of successful viral culturing attempts (SARS-CoV-2)	Day 14
Viral culturing - Day 17	Number of successful viral culturing attempts (SARS-CoV-2)	Day 17
Viral culturing - Day 24	Number of successful viral culturing attempts (SARS-CoV-2)	Day 24
Viral culturing - Day 30	Number of successful viral culturing attempts (SARS-CoV-2)	Day 30

Secondary Outcome Measures

Outcome Measure	Measure Description	Time Frame
Baseline Viral Load - Day 0	Log10 copies/ml	Day 0
Change from Baseline Viral Load - Day 3	Log10 copies/ml	Day 3
Change from Baseline Viral Load - Day 7	Log10 copies/ml	Day 7
Change from Baseline Viral Load - Day 10	Log10 copies/ml	Day 10
Change from Baseline Viral Load - Day 14	Log10 copies/ml	Day 14
Change from Baseline Viral Load - Day 17	Log10 copies/ml	Day 17
Change from Baseline Viral Load - Day 24	Log10 copies/ml	Day 24
Change from Baseline Viral Load - Day 30	Log10 copies/ml	Day 30

Collaborators and Investigators

• Sponsor: Nordsjaellands Hospital
• Collaborators: Rigshospitalet, Denmark; Statens Serum Institut
• Investigators: Principal Investigator: Zitta Barrella Harboe, MD, PhD, Nordsjaellands Hospital

Study record dates

Study Major Dates

• Study Start (Actual): May 24, 2020
• Primary Completion (Actual): May 5, 2021
• Study Completion (Anticipated): October 1, 2023

Study Registration Dates

• First Submitted: March 8, 2022
• First Submitted That Met QC Criteria: March 9, 2022
• First Posted (Actual): March 10, 2022

Study Record Updates

• Last Update Posted (Actual): March 10, 2022
• Last Update Submitted That Met QC Criteria: March 9, 2022
• Last Verified: March 1, 2022

More Information

Terms related to this study

• Keywords: Viral Load; Neutralizing Antibodies; Antibody Decay; Viral Culturing; Infectiousness
• Additional Relevant MeSH Terms: Coronavirus Infections; Coronaviridae Infections; Nidovirales Infections; RNA Virus Infections; Virus Diseases; Infections; Respiratory Tract Infections; Respiratory Tract Diseases; Pneumonia, Viral; Pneumonia; Lung Diseases; COVID-19
• Other Study ID Numbers: Covisera

Drug and device information, study documents

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