Assessment of the Influence of Clinical, Functional, Immunological, and Genetic Factors on the Severity of the Course of Coronavirus Infection With SARS-CoV-2 and Post Covid Syndrome
Conditions: Covid19, Corona Virus Infection
Sponsor: National Research Center for Cardiac Surgery, Kazakhstan

 
 
Title: Assessment of the influence of 
clinical, 
functional, 
immunological 
and 
genetic factors on the severity of the course 
of coronavirus infection with SARS-CoV-2 
and Post Covid syndrome 
 
 
 
NCT number: BR10965164 
 
 
 
Date: 19.04.2021 
 
 

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1. Annotation 
The first case of pneumonia of unknown origin was registered in Wuhan, China in December 
2019 [1]. Shortly after the first case was discovered, the disease was reported outside of China. On 
January 30, 2020, the World Health Organization (WHO) announced the disease was inscribed on the 
International Health Emergencies List (PHEIC), which meant that it posed a threat to the entire world and 
required a coordinated international emergency response [1, 2]. Already on February 11, 2020, the World 
Health Organization assigned the name COVID-19 to the disease [1]. On the same day, the International 
Commission on Virus Classification announced that the new coronavirus had been classified as type 2 
coronavirus, severe acute respiratory syndrome (SARS-CoV-2) [1]. The rapid spread of this infection in 
the first months of 2020 reached pandemic proportions. To date, more than 100 million cases of COVID-
19 and more than 2 million deaths have been recorded in the world [3]. 
COVID-19 is not the first outbreak of severe respiratory illness caused by a virus from the 
coronavirus family. In the past two decades alone, coronaviruses have caused three outbreaks: COVID-
19, Severe Acute Respiratory Syndrome (SARS) and Middle East Respiratory Syndrome (MERS) [4]. 
Although the mortality rate from SARS-CoV-2 is lower than from two previous outbreaks of coronavirus, 
its lightning-fast spread, which led to the development of a pandemic, has caused catastrophic 
consequences for the world. That is why a quick and comprehensive study of the pathogenesis of SARS-
CoV-2 is so important. 
Currently, the world is actively studying the immune pathogenesis of COVID-19, the search for 
measures to prevent and develop drugs against SARS-CoV-2 infection based on previous progress in 
SARS-CoV and MERS-CoV research. Interestingly, COVID-19 can cause a wide range of diseases, 
ranging from asymptomatic to severe respiratory distress syndrome and respiratory failure. However, the 
role of humoral and cellular immunity in the mechanism of the “cytokine storm” and various factors 
leading to a high incidence of complications and mortality from COVID-19 is still not fully understood. 
According to a report in the Lancet, Acute Respiratory Distress Syndrome (ARDS) is the main 
cause of mortality in COVID-19, in the mechanism of which a cytokine storm plays an important role - a 
fatal uncontrolled systemic inflammatory response resulting from the release of large amounts of pro-
inflammatory cytokines and chemokines [5, 6]. That is why, often in the treatment of COVID-19, the use 
of antiviral agents is not enough and it is necessary to carry out complex treatment with steroids and other 
medications aimed at suppressing the immune response [6,7]. There is already evidence that the 
development of a cytokine storm, and, accordingly, respiratory complications is largely associated with 
genetic factors [7]. However, in order to determine the exact data, it is necessary to carry out additional, 
complex research. 
2. Explanatory note 

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 1. General information 
1.1. Name of the topic of the scientific, scientific and technical program: "Assessment of the 
influence of clinical, functional, immunological and genetic factors on the severity of the course of 
coronavirus infection with SARS-CoV-2 and Post Covid syndrome" 
1.2. A strategically important state task for the solution of which a program has been developed - the 
Message of El Basy dated January 10, 2018 "New development opportunities in the context of the fourth 
industrial revolution", in terms of the development of personalized medicine; Clause 5.5 Improving the 
provision of medical care of the State Program for the Development of Healthcare of the Republic of 
Kazakhstan for 2020 - 2025: 
Measures to combat the main diseases leading to mortality are diseases of the circulatory system 
(strokes, heart attacks and others), respiratory diseases, oncological diseases, injuries, health care for 
children and mothers, neurological diseases, and in other clinical services includes: transfer and 
introduction of new and innovative technologies for the diagnosis and treatment of diseases within the 
framework of public and private initiatives 
1.3. Place of implementation of the program: NJSC "National Research Center for Cardiac Surgery" 
1.4. Estimated start and end date of the program, its duration in months: 2021 - 2023, 36 months. 
1.5. Organization-applicant of the program: NJSC "National Research Center for Cardiac Surgery" 
1.6. Executors of the program: 
Private institution "National Laboratory Astana", Nazarbayev University 
School of Medicine, Nazarbayev University 
Joint Stock Company "National Center for Neurosurgery" 
1.8. Key words characterizing the industry and the direction of the program for the selection of 
independent experts: Coronavirus infection, Post Covid syndrome, COVID-19, pneumonia, cardiology, 
genetic research, immunology. 
  2. The general concept of the program. 
2.1. Introductory part. 
Principal Investigator - Bekbossynova Makhabbat, Doctor of Medical Sciences, Acting Chairman 
of the Board of the NJSC "National Research Center for Cardiac Surgery ". 
Responsible person - Tauekelova Ainur Tulepbergenovna, cardiologist, head of the admission 
department, NJSC "National Research Center for Cardiac Surgery". 
On the basis of the NJSC "National Research Center for Cardiac Surgery" an infectious hospital has 
been deployed in June 2020 for hospitalization of persons with confirmed cases of severe coronavirus 
infection. By order of the Ministry of Health of the Republic of Kazakhstan No. 763 dated November 24, 
2020, the country's first postcovid center was created. In December 2020, on the basis of the postcovid 

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center, 312 adult patients and 100 children were examined after suffering a coronavirus infection with a 
confirmed COVID-19 diagnosis. 
2. 2. 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. 
2.3. Objectives of the program: 
1. 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. 
2. To study the immunological profile of patients with varying degrees of the course of the acute 
phase of COVID-19 and Post Covid syndrome. 
3. To study the genetic profile of patients with varying degrees of the course of the acute phase of 
COVID-19 and Post Covid syndrome.  
4. Identify potential predictors of COVID-19 severity.  
5. To determine the markers that allows predicting the development of the Post Covid syndrome.  
6. Based on the selected markers, develop a COVID-19 outcome scale to determine the tactics of 
patient management to prevent the development of Post Covid syndrome. 
3. Scientific innovativnes and significance of the program. 
Immunological research has largely become central to the study of the pathogenesis of COVID-
19. Today, all aspects of the immune system are being studied in the world in order to better understand 
the causes of complications and, possibly, to develop targeted treatments to avoid them. The recent 
publication in the journal Immunology was the first work that helped to systematize scientific data on this 
issue [8]. According to this study, an important feature of COVID-19 is its ability to evade an innate 
immune response [8]. Thus, in patients with a severe form of coronavirus, a significant violation of the 
expression of interferon (IFN-I), which plays an important role for antiviral protection, was noted in 
relation to patients with moderate and mild forms [8]. Noted, 
Another feature of SARS-CoV-2 is its complex effect on all the germs of the immune system. 
Many studies have shown that coronavirus can lead to a decrease in natural killer (NK) levels, which also 
correlates with the severity of the disease [8]. In a more detailed study of the role of NK cells in the 
pathogenesis of COVID-19, it was suggested that their stimulation can help eliminate infection and 

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prevent the development of ARDS [8]. In addition to NK cells, studies are continuing on the role of the 
T-cell response in patients with coronavirus infection. It was noted that lymphopenia (especially a 
decrease in the level of CD8 T cells) can serve as a prognostic factor indicating a more severe course of 
the disease [8]. Moreover, the authors note that a violation of the activity of T cells can lead to 
immunopathology of the lungs and, therefore, to the development of complications in patients with 
COVID-19 [8]. 
Humoral immunity undoubtedly plays a key role in neutralizing the virus. In disease, antibodies 
are usually found that bind the internal N-protein SARS-CoV-2 and the external glycoprotein S [8]. The 
receptor binding domain (RBD) of protein S is highly immunogenic, and antibodies that bind this domain 
are likely to have a high neutralizing ability to block COVID-19 when it binds to human cells [8]. An 
important issue in the study of the consequences of coronavirus infection is the identification of immunity 
after an illness. It is noted that B-cell memory is key in this issue, however, it is still not clear how long it 
can persist in the human body. Long-term protection is achieved through the production of long-lived 
plasma cells and memory B-cells, but the life cycle of these cells after suffering COVID-19 is still not 
fully understood [8]. 
There are also suggestions about the role of antigen presenting cells (APCs) in the pathogenesis of 
COVID-19. They are important in the development of antiviral immunity in the body and are recognized 
by virus-specific cytotoxic T-lymphocytes (CTL) [9]. Today it is becoming clear that COVID-19 can 
cause a complex systemic reaction between various components of the immune system [10]. Therefore, 
additional research on this topic will help us understand the pathogenesis of COVID-19 and the role of 
humoral and cellular immunity in the development of complications. 
There are still controversial issues in the diagnosis of COVID-19. To date, the standard for 
diagnosis of COVID-19 is reverse transcription polymerase chain reaction (PCR), which includes the 
detection of viral nucleic acid in sputum or nasopharyngeal swab. But as practice has shown, this test has 
several drawbacks, ranging from expensive cost to low sensitivity, which can affect a truly positive result. 
In addition, the occurrence of conflicting results between chest CT and PCR, described in some studies, 
lead us to the conclusion that the results of the PCR test should always be interpreted in a broader context, 
taking into account other diagnostic markers and the clinical picture [11,12]. 
Various complications of COVID-19 have become one of the significant and dangerous outcomes 
of coronavirus infections. Respiratory failure caused by pneumonia, acute respiratory distress syndrome, 
diabetes mellitus, severe heart damage with a significant increase in the level of troponin in the blood and 
heart failure became the causes of high mortality. 
In particular, the reported mortality rates for COVID-19 are 10.5% in patients with cardiovascular 
diseases, 7.3% in patients with diabetes and 6.0% in patients with arterial hypertension [13]. This is 
higher than the mortality rate observed globally in patients without concomitant diseases, which is 3-4% 

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[13]. Finally, the increased incidence of unwanted cardiovascular and other complications after recovery 
from COVID-19 may also play a role in the increased mortality rate of patients with coronavirus 
infection. 
Although there is still no officially clinically proven specific antiviral agent for the treatment of 
SARS-CoV-2 in the world, symptomatic treatment, including oxygen therapy, anticoagulants, and the use 
of broad-spectrum antibiotics for the treatment of secondary complications, remains important in the 
treatment strategy [14]. 
It is not yet known whether SARS-CoV-2 infection leads to sustained protective immunity, by 
what mechanism this can occur, and whether the vaccine-induced immune responses will protect without 
causing harm [5]. According to studies of the molecular mechanisms of the SARS-CoV-2 genomes, there 
are currently several potential targets for effective therapy, but nevertheless, there are more and more 
open questions requiring detailed study [15,16]. 
There is already evidence that COVID-19 can lead to long-term consequences and exacerbation of 
chronic diseases [17, 18]. The so-called longcovid or ongoing symptomatic COVID-19 is characterized 
by symptoms lasting from 3 to 12 weeks after suffering COVID-19 [18,19]. If complications last more 
than 12 weeks, the syndrome becomes postquoid [19]. Symptoms vary greatly from person to person and 
further study is necessary to develop management strategies for these patients. The world community has 
already noted that further study of long and postcovid syndromes is necessary to prevent the development 
of even greater disorders [20]. 
As of today, the WHO has updated the ICB-10 database on COVID-19 coding. Code U09.9 now 
encodes "Condition after COVID-19", which includes loncoid and Post Covid syndromes [21]. However, 
the timing of the course of this pathology, symptoms, risk factors and other data have not yet been 
determined and need more detailed study. The study of the consequences of the postponed coronavirus 
infection continues in order to predict the epidemiological situation in the world. The situation is 
aggravated by the fact that the course, severity of the disease and complications in patients are very 
different. Thus, studying the genetic epidemiology of patients with COVID-19 in a complex of factors 
influencing the course of the disease and outcomes will help develop measures to combat the epidemic of 
the virus. 
Thus, the above data served as the purpose of this research work, which for the first time in 
Kazakhstan will become work on the study of pathogenetic mechanisms, immunological and genetic 
markers for the development of effective treatment methods, determination of prognostic factors and 
management of patients with COVID-19 and Post Covid syndrome to prevent the development of 
subsequent complications. 

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For the first time, a comprehensive assessment of the severity of the course and outcomes of 
coronavirus infection will be carried out in the context of clinical, functional, immunological and genetic 
factors:  
1. The relationship between the immune response, determined by genetic status, and the severity of 
the course in patients with coronavirus infection will be investigated  
2. Risk factors for the development of long / Post Covid syndromes will be identified.  
3. Genetic characteristics of patients with long / Post Covid syndrome were determined to identify 
potential predictors of severity, predict the course and outcomes of the disease. 
4. The immunological profile of patients after coronavirus infection will be studied in dynamics to 
assess its role in the development of long / Post Covid syndromes.  
The identified predictors will be used to develop a scale of COVID-19 outcomes for predicting and 
determining various course options, which will allow introducing a treatment algorithm to prevent the 
development of Post Covid syndrome, complications and disability in patients who have undergone 
coronavirus infection.  
4. Research methods and ethical issues. 
This study is multicenter, cohort controlled, comparative and prospective. The selection of 
patients will be carried out among patients who have undergone COVID-19 admitted to the National 
Research Center for Cardiac Surgery and  Republican Diagnostic Center of the University Medical Center 
corporate fund using laboratory-functional tests in accordance with standard clinical practice. 
The objects of the study will be: patients who have undergone COVID-19.  
Sample size 300 patients 
Materials and methods: 
Inclusion criteria 
 Age over 18 years old 
 Patients with a history of coronavirus infection COVID-19 confirmed by PCR analysis  
 Patients who signed informed consent to participate in the study 
 
 
Exclusion criteria: 
 Refusal to undergo diagnostic procedures determined by the research protocol. 

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 Evidence for preexisting interstitial lung disease.  
 Participation in another study. 
 
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. 
Further, on the basis of complex data, the severity of the course of the disease will be determined, 
followed by distribution and analysis (Figure 1).  
 
Figure 1. Dividing patients into groups 
 
For the study, clinical-functional, immunological and genetic research methods will be used in 
accordance with the republican and international recommendations for the treatment of patients with 

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COVID-19.  
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 
 EchoCG + strain 
 CT scan of the lungs 
 Holter 
 SMAD 
 Kidney ultrasound 
 Ultrasound  
 Doppler ultrasonography of veins and arteries 
 Chalder Scale, EQ Questionnaire 
Table 1. Clinical and functional analysis 
 
Neurological disorders: 

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1. 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:  
a. The study of antibodies to some neurospecific antigens - myelin basic protein (MBP), 
neurospecific enolase (NSE). 
b. Study of cellular immunity (CD3 +, CD4 +, CD8 +) and general indicators of humoral 
immunity (IgG, IgA, IgM, circulating immune complexes). 
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 

11 
 
 
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. 
 

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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. 
 
 
Figure 2. Frequency of visits and sample collection (in months) 
Visits 
Month 
0 
1 
2 
3 
4 
5 
6 
7 
8 
9 
10 
11 
12 
Analysis 
Visit - doctor's consultation 
1 
1 
1 
1 
1 
1 
1 
1 
1 
1 
1 
1 
1 
Written agreement 
1 
  
  
  
  
  
  
  
  
  
  
  
  
Blood tests: KLA, Neutrophil to 
lymphocyte ratio 
1 
1 
1 
  
  
  
1 
  
  
  
  
  
1 
Blood type 
1 
  
  
  
  
  
  
  
  
  
  
  
  
ALT, AST, total and direct 
bilirubin, LDH, CRP, alpha-
amylase, creatinine, urea, 
glucose,  
1 
1 
1 
  
  
  
1 
  
  
  
  
  
1 
Ferritin 
1 
1 
1 
  
  
  
1 
  
  
  
  
  
1  
Glycohemoglobin 
1 
  
  
  
  
  
1 
  
  
  
  
  
1 
 25 - OH vitamin D 
1 
  
  
  
  
  
1 
  
  
  
  
  
1 
vitamin B12 
1 
  
  
  
  
  
1 
  
  
  
  
  
  

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D-dimer, fibrinogen, INR, APTT 
1 
1 
1 
  
  
  
1 
  
  
  
  
  
1 
ICL IgM / IgG, RBD, S1, N 
1 
1 
1 
1 
1 
1 
1 
1 
1 
1 
1 
1 
1 
NT - pro BNP 
1 
  
  
  
  
  
1 
  
  
  
  
  
1 
Homocysteine 
1 
  
  
  
  
  
1 
  
  
  
  
  
1 
Troponin 
1 
  
  
  
  
  
  
  
  
  
  
  
  
HR, NPV, BP, t C, Saturation 
1 
1 
1 
1 
1 
1 
1 
1 
1 
1 
1 
1 
1 
ECG 
1 
1 
  
  
  
  
1 
  
  
  
  
  
1 
EchoCG + Strain 
1 
  
  
  
  
  
1 
  
  
  
  
  
1 
CT Abdominal 
1 
  
  
1 
  
  
  
  
  
  
  
  
1 
HMECG 
1 
  
  
  
  
  
  
  
  
  
  
  
  
SMAD 
1 
  
  
  
  
  
  
  
  
  
  
  
  
Abdominal ultrasound  
1 
  
  
  
  
  
  
  
  
  
  
  
  
Ultrasound of the kidneys 
1 
  
  
  
  
  
  
  
  
  
  
  
  
Doppler ultrasonography of veins 
and arteries 
1 
  
  
1 
  
  
  
  
  
  
  
  
  
Antibodies to myelin basic 
protein (MBP), neurospecific 
enolase (NSE)  
1 
  
  
  
  
  
  
  
  
  
  
  
  
EEG, CT - perfusion, 
polysonography 
1 
  
  
  
  
  
  
  
  
  
  
  
  
Genetic analysis 
1 
  
  
  
  
  
  
  
  
  
  
  
  
Evaluation of New LongCovid 
Symptoms, PostCovid 
1 
1 
1 
1 
1 
1 
1 
1 
1 
1 
1 
1 
1 
6-minute walk test 
1 
  
  
  
  
  
1 
  
  
  
  
  
1 
Chadler's questionnaire 
1 
1 
1 
1 
1 
1 
1 
1 
1 
1 
1 
1 
1 
EQ questionnaire 
1 
  
  
1 
  
  
1 
  
  
  
  
  
1 
Anxiety and Depression Scale 
1 
1 
1 
1 
  
  
1 
  
  
  
  
  
1 
Summary Mental Status 
Evaluation (MMSE) 
1 
  
  
  
  
  
1 
  
  
  
  
  
1 
 
Immunofetnotyping of a 
subpopulation of T and B 
lymphocytes CD3, CD4, CD5, CD8, 
CD19, CD27, CD38, IgD, anti-
CCR7, anti-CD45RO, NK - cells 
 
1 
1 
1 
1 
1 
1 
Multiplex immunoassay 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, TNF-β, 
VEGF, IgA, IgG1-G4, IgM. 
 
 
 
 
 
 

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Research materials: 
1) blood 
2) deoxyribonucleic acid (DNA) samples 
The study will be conducted in accordance with international standards GCP (Good Clinical Practice) 
in compliance with the principles of scientific ethics, as well as in accordance with the Order of the 
Minister of Health of the Republic of Kazakhstan dated December 14, 2020 No. 248 "On approval of the 
Rules for conducting biomedical experiments, preclinical (non-clinical) and clinical trials , as well as 
requirements for preclinical and clinical bases ”. 
Patient safety procedures will be strictly followed in accordance with the requirements of the Joint 
Commission International (JCI). Each patient will be introduced to the nature of the study. The doctor - 
researcher will explain in detail the essence, benefits and risks of this study. If the decision is positive, the 
patient will sign an informed consent for this study. 
 
The main composition of the research group: 
The qualification of the program group allows for extensive and versatile systematic research in 
the chosen direction. In addition, the program participants have sufficient work experience and 
professional qualifications, which allows obtaining reliable data using modern equipment. The project 
executors have repeatedly participated in competitions. 
 
NJSC "National Research Center for Cardiac Surgery" 
1. Tauekelova Ainur - head of the admission department of the National Research Center for 
Cardiac Surgery 
2. Shestak Elena - cardiologist, National Research Center for Cardiac Surgery. 
3. Dalbekova Marzhan Kairatovna - cardiologist, National Research Center for Cardiac Surgery. 
4. Seitkasym Sholpan Kanatkyzy - cardiologist, National Research Center for Cardiac Surgery. 
5. Litvinova Lia - Clinical Pharmacologist, National Research Center for Cardiac Surgery. 
6. Ivanova-Razumova Tatyana - Cardiologist, National Research Center for Cardiac Surgery  
7. Saylybayeva Aliya Ibaydullaevna, PhD, Head of Science Department, National Research Center 
for Cardiac Surgery 
8. Daniyarova Gulnur - Scientific Secretary, National Research Center for Cardiac Surgery  
9. Khamitov Sadyk - Chief Specialist of the Science Department, National Research Center for 
Cardiac Surgery 
10. Vacancy - accountant of National Research Center for Cardiac Surgery 
11. Vacancy - lawyer of the National Research Center for Cardiac Surgery 

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12. Ayapbergenova Roza - economist, National Research Center for Cardiac Surgery 
13. Vacancy - resident doctor of National Research Center for Cardiac Surgery  
14. Vacancy - resident doctor of NJSC National Research Center for Cardiac Surgery 
 
School of Medicine, Nazarbayev University 
15. Dimitri Poddige, Associate Professor of the Faculty of Medicine of Nazarbayev University, 
Director of the Clinical Academic Department of Pediatrics at the University Medical Center 
(UMC). 
From 2007 to 2009, Dr. Dimitri Poddige worked as an Immunology Research Fellow at the Boston 
Children's Hospital at Harvard University in Boston, Massachusetts, USA. Since 2010, Dr. Dimitri 
Poddige has worked as a general practitioner in the pediatric departments of several hospitals in  
Lombardy (Italy), including the Fondazione IRCCS Policlinico San Matteo (Pavia), in collaboration 
with the University of Pavia, where he worked as a teacher / instructor in the Department of 
Pediatrics since 2015. 
 
16. Kuanysh Dosybaeva, M.Sc. Molecular Medicine, Research Assistant, School of Medicine, 
Nazarbayev University. 
 
17. Vacancy, Doctoral student, School of Medicine Nazarbayev University. 
 
Joint Stock Company "National Center for Neurosurgery" 
 
18. Jamantaeva Botagoz Daukimovna, JSC " NCN " Head of the Department of Neurosurgery and 
Neuropathology.  
 
19. Sakhipova Aiym Garipullayevna, JSC "NCN", Chief specialist of the Republican Coordination 
Center for Stroke Problems.  
 
20. Guldana Zhumabayeva, JSC "NCN", leading specialist of the Republican Coordination Center 
for Stroke Problems. 
 
 
6. Research environment. 
NJSC "National Research Center for Cardiac Surgery" has a sufficient material and technical base. 

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Participating organizations are equipped with modern equipment and apparatus for conducting 
clinical and laboratory, instrumental and specific methods according to the profile. The declared 
equipment for research is available and has passed the technical inspection with calibration. Also, there 
are standardized protocols for research methods. 
The program will involve young scientists, doctoral students, undergraduates and residents. 
The program is carried out on a permanent basis at the National Research Center for Cardiac 
Surgery, while molecular genetic methods and specific diagnostic methods will be carried out in profiling 
laboratories. As the temporary full-time team of each organization becomes busy, it is planned that 
employees will participate in scientific work on general tasks and sections, which will increase efficiency 
and reduce work time. 
Laboratory of Genomic and Personalized Medicine and the Laboratory of Bioinformatics and 
Systems Biology of the Center for Life Sciences have the necessary equipment, material and technical 
base, equipment that is standard for world-class genomic laboratories, as well as necessary for high-
throughput sequencing. For analysis of full genome sequencing data and storage of information, there are 
servers and computer stations. For storage of biological samples, there are freezers -80 C. The list of the 
main units of equipment and technology is given in the table, except for laboratory small-sized 
equipment, which is necessary for the project. 
 
9. Expected results of the program. 
Patients who have undergone a coronavirus infection and have received a number of 
complications that affect the quality and duration of life are one of the urgent problems in the world. The 
situation is aggravated by the fact that the issue is still not fully understood due to the novelty of the 
disease, which poses an even greater serious threat to humanity. 
Nevertheless, understanding and studying the problem plays a decisive role not only in health 
care, but in general will affect the economic, social consequences in the world, as well as its safety. 
The implementation of this project, for the first time in the Republic of Kazakhstan, will allow 
studying the main mechanisms of infection, its consequences not only for the body, but also for society as 
a whole.  
The results of this project will contribute to improving public awareness of the causes, prevention 
of infection and complications of coronavirus infection. Physicians in clinical practice will be able to rely 
on the developed criteria for diagnosis and treatment. Medical staff will receive a more detailed picture of 
the most acute and important measures for the management and care of patients with coronavirus 
infection to prevent the frequency of complications and the development of post-ovarian syndrome, and 
will also be able to provide appropriate assistance to patients to adapt to a new lifestyle. 

17 
 
 
In addition, the results of the proposed study will influence the further development of research 
initiatives such as the development of preventive vaccinations, the development of new drugs and the 
study and implementation of new strategies in the treatment of coronavirus complications.  
Along with this, the developed and implemented scale of infection outcomes will significantly 
improve the prognosis and survival of this category of patients and affect the epidemiological situation. 
The target consumers of the results obtained will be patients who have undergone COVID-19, as 
well as the entire population of the Republic of Kazakhstan, which are currently the main value of the 
country.  
At least 2 (two) articles and (or) reviews will be published in peer-reviewed scientific journals in 
the scientific direction of the program, included in 1 (first), 2 (second) or 3 (third) quartiles in the Web of 
Science database and (or) having percentile by CiteScore in the Scopus database not less than 50 (fifty). 
To popularize science, a separate website will be created, which will provide complete information on the 
progress of the program. For each scientific publication within the framework of the program, 
information will be published on the website and / or in social networks. 
In general, the expected efficiency and effectiveness of the proposed project will make a 
significant contribution to the development of not only health care, but will also have a positive 
contribution to the country's economy as a whole. In addition, the effective distribution of budgetary will 
maximize the concentration of the expenditure side of the implementation of this project to achieve the 
designated goals and objectives. 
This project is aimed at achieving the indicators of the State Program for Healthcare Development 
2020-2025. an increase in average life expectancy up to 75 years and a decrease in the risk of premature 
mortality from 30 to 70 years from cardiovascular diseases. 
 
 
 
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