An observational study in Covid19 and Corona Virus Infection, sponsored by National Research Center for Cardiac Surgery, Kazakhstan. Status unknown at 1 site in Kazakhstan. Open to participants aged 18 Years to 65 Years. Per ClinicalTrials.gov, last updated 2021-08-03.
Sponsored by National Research Center for Cardiac Surgery, Kazakhstan · Observational
The purpose of the program. To determine the clinical, functional, immunological, and genetic factors affecting the severity of the course of acute coronavirus infection COVID-19 and PostCovid syndrome, in order to develop management tactics for such patients to reduce the risk of complications and disability.
Objectives of the program:
To determine the clinical and functional characteristics of patients with varying degrees of the course of the acute phase of COVID-19 and Post Covid syndrome.
1.1. To study the features of neurological disorders in patients with varying degrees of the course of the acute phase of COVID-19 and Post Covid syndrome.
The study will include patients with a positive PCR test for COVID-19. Patients in the acute phase of the course of the disease are monitored and treated in accordance with the republican COVID-19 treatment protocol. After signing the informed consent, the patient will be included in the study. The collection of the necessary materials for subsequent analyzes (clinical-functional, genetic, immunological) will be carried out in accordance with this protocol. Subsequently, patients are observed within one year from the moment of illness in accordance with the study protocol and with the collection of all necessary materials.
Clinical and functional analysis:
Detection of RNA of the COVID-19 virus using PCR analysis. Conducting complex laboratory studies in accordance with table 1.
General blood analysis Complete blood count on an analyzer with differentiation of 5 classes of cells, the ratio of neutrophils to lymphocytes
Blood chemistry ALT, AST, total bilirubin, direct, LDH, CRP, alpha-amylase, creatinine, urea, glucose, ferritin, glycosylated hemoglobin, vitamin 25 - OH vitamin D, vitamin B12
Coagulogram D-dimers, fibrinogen, INR, APTT
Other NT-pro BNP, Homocysteine, IL 6, Troponin, blood group determination
Linked immunosorbent assay Determination of IgG and IgM antibodies to SARS-CoV-2 coronavirus (COVID-19) in blood serum, RBD
Functional diagnostics • ECG
Neurological disorders:
Laboratory research methods:
Immunological analysis:
A comprehensive immunological analysis will be carried out to determine the level of the immune response. Calculation of the level of CD4 +, CD8 + and NK cells. The level of antibodies of the IgG and IgM classes to the proteins of the coronavirus S1, RBD and N was determined Multiplex Immunoassay. For evaluation of immunological parameters, samples are diluted in 200 μl of phosphate buffer, centrifuged and the supernatant analyzed using the manufacturer's protocol. The MILLIPLEX MAP human cytokine / chemokine magnetic bead panel will be used for the analysis of multiple cytokines and chemokines / immunoglobulins, and the Milliplex® magnetic bead panel (HGAMMAG-301K-06, EMD Millipore Corp., Billerica, MA) will be used for immunoglobulin isotyping. Samples will be analyzed on Bioplex BIO-RAD for the following indicators: sCD40L, EGF, Eotaxin / CCL11, FGF-2, Flt-3 ligand, Fractalkine, G-CSF, GM-CSF, GRO, IFN-α2, IFN-γ, IL -1α, IL-1β, IL-1ra, IL-2, IL-3, IL-4, IL-5, IL-6, IL-7, IL-8, IL-9, IL-10, IL-12 (p40), IL-12 (p70), IL-13, IL-15, IL-17A, IP-10, MCP-1, MCP-3, MDC (CCL22), MIP-1α, MIP-1β, PDGF- AA, PDGF-AB / BB, RANTES, TGF-α, TNF-α, Immunophenotyping of T-cell and B-cell subpopulations using the flow cytometry method. Subpopulations of B- and T-lymphocytes will be examined by staining peripheral blood mononuclear cells (PBMCs) isolated from whole peripheral blood with monoclonal antibodies conjugated to fluorochromes. PBMCs can be isolated from EDTA-treated whole blood using Ficoll density gradient centrifugation or special erythrocyte lysis buffers. It is preferable to analyze isolated PBMCs directly on the day of blood collection, which gives more reliable results. PBMC viability will be assessed using LIVE / DEAD ™ fixed dead cell kits or 0.4% trypan blue and propidium iodide solution. After lysis of erythrocytes and incubation with monoclonal antibodies, the stained cells are resuspended in a staining medium and examined using a MoFlo Astrios flow cytometer (Beckman Coulter, USA). The resulting data will be analyzed using Summit (Beckman Coulter) and FloJo (Tree Star) software.
Forward and side scatter will be used to distinguish the lymphocyte population in addition to the signal from specific fluorochromes. Distribution CD3-, CD5 +, CD19 + (total number of B-lymphocytes), CD5-, CD19 +, CD27 + (memory B-cells), CD19 + CD27- (naive B-cells), CD19 + CD27 + CD38 + IgD - (Class-Switched Memory B-Cells) CD19 + CD27 + CD38 + IgD + (Unswitched Memory B-Cells) will be analyzed on the general lymphocyte population. The distribution of markers CD3, CD4 and CD8 will be analyzed in the pool of T-lymphocytes. To analyze the differentiation status of T cells, cells are additionally stained with anti-CCR7, anti-CD45RO antibodies. Antibodies will be purchased from Invitrogen ™ unless otherwise noted.
Genetic analysis:
Isolation (extraction) of DNA will be performed from whole blood using commercial kits according to the manufacturer's instructions. To analyze a large number of genetic markers, it is planned to carry out genome-wide sequencing followed by analysis of genetic polymorphisms of candidate genes encoding coronavirus receptors and immunological factors.
Sequencing will be performed using high-throughput next generation sequencing platforms Illumina NovaSeq6000 (Illumina), method validation using traditional capillary sequencing - ABI 3730XL ™ DNA Analyzer (Life Technologies), real-time PCR.
Bioinformatic data analysis.Bioinformatics sequencing data will be analyzed. The software packages for bioinformatic analysis of sequencing data (GATK, bwa, bowtie, bowtie2, VarScan etc) will be used. The sequencing data will be compared with the publicly available data from the world's international databases of genomic research (https://www.covid19hg.org/, ExAC, HGMD (Human Gene Mutation Database), ESP, GeneBank, NCBI, ESP6500, 1000Genomes, SNPDb130, Ensembl, ClinVar, SNPedia, etc.). Differences in the type and frequency of genomic variation among the surveyed groups will be determined.
To classify the detected genetic variants, in silico models will be used (SIFT_score / pred, Polyphen2_HDIVscore / pred, Polyphen2_HVAR_score / pred, LRT_score / pred, MutationTaster_score / pred, MutationAssessor_score / pred, FATHMM_score / pred, Radial / MetaRVM_score pred). The classification of clinically significant genetic variants will be carried out according to the international ACMG / AMG criteria.
Statistical analysis:
Statistical analysis will be carried out using version R 3.6.2. Quantitative data, including clinical, biochemical, molecular genetic parameters, will be checked for normality using the Shapiro-Wilks test and recognized as parametric in distribution. Comparison of mean differences will be performed using one-way ANOVA, and subsequent pairwise comparison will be performed using Tukey's special test. Within-group mean differences will be performed using the paired sample t-test. The graphs will be executed using the ggplot2 R package. Statistical analysis will be performed for the multiplex analysis results using the R psych package and standard t-tests.
Inspection frequency:
Patients are followed up for 12 months from the date of illness. Disease detection corresponds to the baseline (day 0). At the time of diagnosis, materials are taken for clinical, functional and immunological diagnostics. After that, the sampling is carried out every month for the next year from the moment of the disease in accordance with the study protocol (Figure 2). The collection of materials for genetic analysis is carried out once.
7,990 studies on the registry are indexed under COVID-19; 466 are open to participants now.
This study's planned enrollment of 300 is above the median of 268 across 3,208 observational studies indexed under COVID-19.
Browse COVID-19 studies →National Research Center for Cardiac Surgery, Kazakhstan is the lead sponsor of 5 studies on the registry; 1 is open to participants now.
Counted across the registry records on this site, refreshed daily.
The study will include patients with a positive PCR test for COVID-19. Patients in the acute phase of the course of the disease are monitored and treated in accordance with the republican COVID-19 treatment protocol. After signing the informed consent, the patient will be included in the study. The collection of the necessary materials for subsequent analyzes (clinical-functional, genetic, immunological) will be carried out in accordance with this protocol. Subsequently, patients are observed within one year from the moment of illness in accordance with the study protocol and with the collection of all necessary materials.
Go through the criteria one by one — yes, no or not sure — and take the list to your doctor or the study team. Kept in this browser only.
Exclusion Criteria:
Underlined terms in the criteria open these too. Explained by clinicaltrials.gg, not part of the registry record. The study team decides who can take part.
Before you join, the study team explains what taking part involves — its purpose, what happens, the risks and possible benefits — and answers your questions, so you can decide freely. Consent is usually recorded on a form (the informed consent form, ICF) signed by you or, where the law allows, by someone legally authorised to decide for you.
You can keep asking questions during the study, you are told of new information that matters, and you can choose to leave at any time.
Informed consent
Before you join, the study team explains what taking part involves — its purpose, what happens, the risks and possible benefits — and answers your questions, so you can decide freely. Consent is usually recorded on a form (the informed consent form, ICF) signed by you or, where the law allows, by someone legally authorised to decide for you.
You can keep asking questions during the study, you are told of new information that matters, and you can choose to leave at any time.
Explained by clinicaltrials.gg, not part of the registry record. The study team decides who can take part.
Patients in the acute phase of the course of the disease
Diagnostic Test: Clinical and functional analysis · Diagnostic Test: Neurological analysis · Diagnostic Test: Immunological analysis: · Diagnostic Test: Genetic analysis:
Patients with chronic symptoms after a previous Covid-19 (4-12 weeks)
Diagnostic Test: Clinical and functional analysis · Diagnostic Test: Neurological analysis · Diagnostic Test: Immunological analysis: · Diagnostic Test: Genetic analysis:
Patients with chronic symptoms after a previous Covid-19 (more than 12 weeks)
Diagnostic Test: Clinical and functional analysis · Diagnostic Test: Neurological analysis · Diagnostic Test: Immunological analysis: · Diagnostic Test: Genetic analysis:
General blood analysis - Complete blood count on an analyzer with differentiation of 5 classes of cells, the ratio of neutrophils to lymphocytes; Blood chemistry - ALT, AST, total bilirubin, direct, LDH, CRP, alpha-amylase, creatinine, urea, glucose, ferritin, glycosylated hemoglobin, vitamin 25 - OH vitamin D, vitamin B12; Coagulogram - D-dimers, fibrinogen, INR, APTT; Other - NT-pro BNP, Homocysteine, IL 6, Troponin, blood group determination; Linked immunosorbent assay - Determination of IgG and IgM antibodies to SARS-CoV-2 coronavirus (COVID-19) in blood serum, RBD; Functional diagnostics • ECG * EchoCG + strain * CT scan of the lungs * Holter * SMAD * Kidney ultrasound * Ultrasound * Doppler ultrasonography of veins and arteries * Chalder Scale, EQ Questionnaire
Also known as: Immunological analysis, Genetic analysis
Neurological examination with the isolation of neurological syndromes (motor, cognitive impairments, sleep disorders, asthenic-depressive syndromes, etc.). 2. Neuropsychological methods - research on the scales of anxiety and depression, MMSE, etc. 3. Instrumental method - EEG, polysonography, ultrasound of the neck vessels, CT perfusion. 4. Laboratory research methods: 1. The study of antibodies to some neurospecific antigens - myelin basic protein (MBP), neurospecific enolase (NSE). 2. Study of cellular immunity (CD3 +, CD4 +, CD8 +) and general indicators of humoral immunity (IgG, IgA, IgM, circulating immune complexes).
A comprehensive immunological analysis will be carried out to determine the level of the immune response. Calculation of the level of CD4 +, CD8 + and NK cells. The level of antibodies of the IgG and IgM classes to the proteins of the coronavirus S1, RBD and N was determined
Also known as: Multiplex Immunoassay, Immunophenotyping of T-cell and B-cell subpopulations
Isolation (extraction) of DNA will be performed from whole blood using commercial kits according to the manufacturer's instructions. To analyze a large number of genetic markers, it is planned to carry out genome-wide sequencing followed by analysis of genetic polymorphisms of candidate genes encoding coronavirus receptors and immunological factors.
bioinformatic data analysis
To analyze a large number of genetic markers, it is planned to carry out genome-wide sequencing followed by analysis of genetic polymorphisms of candidate genes encoding coronavirus receptors and immunological factors
Time frame: 24 months
the level of the immune response
Calculation of the level of CD4 +, CD8 + and NK cells. The level of antibodies of the IgG and IgM classes to the proteins of the coronavirus S1, RBD and N was determined
Time frame: 24 months
PDFs hosted by the registry, ClinicalTrials.gov.
Plan to share: Yes — all IPD that underlie results in a publication
Supporting information: Study protocol, Statistical analysis plan, Informed consent form, Clinical study report, Analytic code
Time frame: starting 6 months after publication
Access criteria: all types of supporting information will be shared with the interested party within the framework of the legislation of the Republic of Kazakhstan
Data sharing details ↗ · heartcenter.kz
Started Jun 2021 (actual). Primary completion is estimated for Dec 2022, 18 months after the start.
Of 1,640 non-industry observational studies under COVID-19, half reached primary completion within 14 months of starting (the median), counting those still running; a quarter within 5 and three quarters within 30. This study's planned 18 months is above that.
Counting every sponsor, half of 1,774 studies reached it within 13 months; a quarter within 5 and three quarters within 29.
How these figures are worked out, and which studies are left out of them.
This study is status unknown, as verified in Jul 2021. You cannot join it, but the record below documents what was studied.
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National Research Center for Cardiac Surgery, Kazakhstan