RU2765686C1 - Method for prediction of risks of severe course of respiratory diseases of viral nature - Google Patents

Abstract

FIELD: molecular biology; genetics.

SUBSTANCE: described is a method for predicting the risks of severe respiratory diseases of a viral nature, which consists in isolating human DNA, determining the DNA sequence in the field of polymorphisms in the human genome: rs1990760 in the IFIH1 gene, rs12252 in the IFITM3 gene, rs1800629 in the TNF gene, rs75603675 in the TMPRSS2 gene, rs7842 in the C3AR1 gene, rs744166 in the STAT3 gene, rs324011 in the STAT6 gene, rs179008 in the TLR7 gene, rs1799864 in the CCR2 gene, rs1898830 in the TLR2 gene, assessing risks by assigning points to the genotype in the area of each polymorphism, summing these points, wherein the total score = "0" is the general population risk, the total score is less than "0" — the risk is increased, the total score is more than "0" — the risk is lowered.

EFFECT: invention can be used for prediction of genetic predisposition of a person to severe or complicated course of respiratory disease of viral nature and increased or reduced predisposition to it.

1 cl, 5 tbl, 3 ex

Description

Technical field

The invention relates to molecular biology and genetics and can be used in virology, immunology and medicine to predict a person's genetic predisposition to a severe or complicated course of a respiratory disease of a viral nature and an increased or decreased predisposition to it.

State of the art

Respiratory diseases of a viral nature are caused by both RNA-containing viruses (families Orthomyxoviridae , Paramyxoviridae , Picobirnaviridae , Picornaviridae , Coronaviridae ) and DNA-containing viruses (families Adenoviridae , Parvoviridae ). These diseases remain the leading cause of death, disability and social and economic unrest for millions of people around the world. Poverty, lack of adequate access to health care, human migration, the emergence of new pathogens and the use of antibiotics for viral infections all contribute to the increased impact of these diseases.

Identification of specific polymorphisms in human genes that are associated with susceptibility or resistance to viral diseases is a promising direction in medical diagnostics. Human knowledge of the genetic factors involved in the pathogenesis of viral infections, and especially in the pathogenesis of their complications, continues to grow. The main goal of genetic research in the field of infectious diseases is to predict susceptibility, prevent complications and timely selected therapy to reduce the severity of the disease (Adam D. Kenney, James A. Dowdle, Leonia Bozzacco, Themet M. McMichael, Corine St. Gelais, Amanda R. Panfil, YanSun, LarryS. Schlesinger, Matthew Z. Anderson, Patrik L. Green, Carolina B. Lopez, Brad R. Rosenberg, LiWuand Jacob S. Yount. HUMAN GENETIC DETERMINANTS OF VIRAL DISEASES.Annu Rev Genet.2017 November 27;51:241-263).

Known solution according to the application USA No. US20160273044 A1, C12Q1/6883, which consists in the identification, diagnosis and prognosis of systemic lupus erythematosus, including treatment methods, built on the basis of point genotyping of polymorphisms in genes associated with immunity, including polymorphism rs1990760 ( IFIH1 ), where T the allele is a risk factor. The IFIH1 gene belongs to the family of pattern recognition receptors, is a sensor of viral nucleic acids in the cytoplasm, and plays a major role in recognizing a viral infection and activating a cascade of antiviral responses: stimulating the release of type I interferons and pro-inflammatory cytokines.

However, this method applies only to autoimmune diseases, and although it affects most of the genes of the signaling pathways of viral pathogenesis, it is not infectious.

Solutions are also known regarding the risk assessment of the severity of an infectious disease based on the determination of various biomarkers.

Application WO 2017/050988 Al describes a method for predicting the severity of influenza based on the detection of a protein marker, interferon-alpha-inducible protein 27.

Also, in the application WO2014/008545A1, the level of the same protein is considered as a diagnostic criterion for viral pneumonia and as a prognostic criterion for the severity of the disease.

The disadvantage of this method is that alpha-interferon-inducible protein 27 is a marker of only one of the signaling pathways of the immune response. Also, the disadvantage is that this protein is detected already at the stage of the development of the disease, and the identification of the risk of complications may occur too late.

Genetic markers, as a risk criterion for the severity of the disease, are promising, since they can be analyzed at any time and at any age, and the choice of therapy according to the genotype can be made at the very beginning of the disease, without additional analyzes.

Some genetic markers of the severity of viral respiratory infection are described in the scientific literature.

For example, in H1N1 infection in 2009, a significantly higher frequency of the TNF rs1800629 G/A genotype was found in severe and fatal cases (ChoudharyM.L., AlagarasuK., ChaudharyU., KawaleS., MalasaneP., GuravY.K., PadbidriV., KadamD., SangleS.A., SalviS., BavdekarA.R., D'costaP., ChadhaM.S.AssociationofSingleNucleotidePolymorphismsinTNFAandIL10 GeneswithDiseaseSeverityinInfluenzaA/H1N1pdm09 VirusInfections: AStudyfromWesternIndia.ViralImmunol.2018 Dec;31(108):683).

A solution is known according to patent RU No. 2339701, which considers a method for determining a person's predisposition to various types of physical work based on a genetic panel that includes a number of polymorphisms. But the method considered in this patent is not suitable for determining predisposition to viral infections, since the genes included in the panel are not associated with the human immune system or with the regulation of the inflammatory process.

A decision is known according to patent RU No. 2494400, which discusses a method for predicting pregnancy pathologies based on the determination of polymorphism in the PAI-1 gene.

But the method considered in this patent is not suitable for determining predisposition to viral infections, since although the PAI-1 gene is associated with the development of an inflammatory process, including in infectious diseases, this polymorphism is not known to be associated with the risk of a respiratory viral infection or with its heaviness.

Disclosure of invention

The objective of the invention is to create a method for predicting the risks of severe viral respiratory diseases in humans and predisposition to them, as well as to create a genetic panel for implementing the method by performing a genetic analysis of several polymorphisms at once.

The technical result is achieved due to the genetic panel created by the authors, which includes polymorphisms that are significantly associated with the risk of a respiratory viral infection or its severe course, which makes it possible to implement a method for predicting the risks of severe respiratory diseases of a viral nature in humans and predisposition to them by conducting genetic analysis. in the field of polymorphisms in the human genome, several polymorphisms in 10 genes at once. In this case, risk assessment is performed by genotype in the area of polymorphisms included in this panel.

The technical task is carried out due to the fact that a genetic panel has been created to predict the risks of severe respiratory diseases of a viral nature and predisposition to them based on genetic research methods, including the genotype in the field of pathogenic or protective alleles of polymorphisms in the human genome: rs1990760 in the IFIH1 gene, rs12252 in The IFITM3 , RS1800629 gene in the TNF gene, RS75603675 in the gene TMPRSS2 , RS7842 in the C3AR1 , RS744166 gene in the STAT3 gene, RS324011 gene in the GENE 6 , RS179008 in the CCR2 gene, RS1898830 gene in the TLR2 gene, while

Gene Polymorphism and genotype in his area Significant genotype Functional value IFIH1 rs1990760; T/C CC, CT Risk IFITM3 rs12252; A/G GG Risk TNF rs1800629; G/A GA, AA Risk TMPRSS2 rs75603675; C/A CA, AA Risk C3AR1 rs7842; T/C TC, CC Risk STAT3 rs744166; A/G AG, GG Protective STAT6 rs324011; C/T CT, TT Risk TLR7 rs179008; A/T TT Risk CCR2 rs1799864; G/A GA, AA Risk TLR2 rs1898830; A/G GG Protective

A method for predicting the risks of severe course of respiratory diseases of a viral nature and predisposition to them has been created, which consists in isolating human DNA, determining the DNA sequence in the field of polymorphisms in the human genome: rs1990760 in the IFIH1 gene, rs12252 in the IFITM3 gene, rs1800629 in the TNF gene, rs75603675 in the TMPRSS2 gene , rs7842 in the C3AR1 gene, rs744166 in the STAT3 gene, rs324011 in the STAT6 gene, rs179008 in the TLR7 gene, rs1799864 in the CCR2 gene, rs1898830 in the TLR2 gene according to claim 1, after which the resulting genotype is analyzed taking into account possible allelic variants of polymorphism. predisposition by summing up the effects of different pathogenic or protective alleles of polymorphisms from claim 1, then the identified genotype is evaluated according to the pathogenic or protective role of alleles in the development of an infectious disease, summarized and evaluated the overall result. either by real-time PCR or by analysis of the length of restriction fragments, and the risks of severe respiratory diseases of a viral nature and predisposition to them are assessed by assigning points to the genotype in the area of each polymorphism, summing these points and assessing risks depending on the sum of points, with In this case, the sum of points = "0" - the risk is general population, the sum of points is less than "0" - the risk is increased, the sum of points is more than "0" - the risk is reduced, and,

genotype - points gene polymorphism homozygous for the major allele heterozygous homozygous for the minor allele CCR2 rs1799864 GG "0" GA "-2" AA "-2" IFIH1 rs1990760 TT "0" TC "0" CC "-2" IFITM3 rs12252 AA "0" AG "0" GG "-3" TNF rs1800629 GG "0" GA "-1" AA "-1" TMPRSS2 rs75603675 CC "0" CA "-1" AA "-1" C3AR1 rs7842 TT "0" TC "-2" CC "-2" STAT3 rs744166 AA "0" AG "0" GG "+2" STAT6 rs324011 CC "0" CT "0" TT "-1" TLR2 rs1898830 AA "0" AG "0" GG "+1" TLR7 rs179008 AA "0" AG "0" GG "-1"

Implementation of the invention

The invention is based on a method for determining a person's predisposition to a respiratory viral infection, as well as to a severe or complicated course of a respiratory viral disease using a genetic panel created by the authors, which allows genetic analysis in the field of polymorphisms in the human genome, several polymorphisms in 10 genes at once.

Improved accuracy, more complete recommendations are based on the introduction of more polymorphisms in more genes and interpretation based on the summation of the effects of the influence of different genotypes as the sample of examined people increases.

In the mechanism of the body's immune response to viruses that cause respiratory infections, the following signaling pathways analyzed by the authors can be distinguished.

The first pathway is initiated by membrane toll-like receptors TLR2 and TLR4, which are pattern-recognizing molecules in relation to pathogens. This pathway ultimately leads to the activation of genes for pro-inflammatory cytokines (interleukin 1-beta, interleukin-6, alpha-TNF) and ensures the development of early inflammatory reactions.

The second signaling pathway begins with the recognition of viral nucleic acids by intracellular toll-like receptors TLR3, TLR7, and TLR8. This pathway leads to the activation of antiviral protection and a late inflammatory response due to the production of interferons alpha and beta.

The third way is the way of activation of the complement system. The genes encoding components of this pathway were also included in the study.

That is, all elements of these signaling pathways play a role in the development of a timely immune response, so mutations in the genes encoding signal chain proteins or regulating the activity of signaling proteins may be responsible for the course of a viral disease.

Antiviral proteins

Transmission of a signal from a pathogen into a cell, activation of the response of the immune system (including the antiviral response) depends on immune modulators, one of which is the product of the IFIH1 gene. This protein is able to elicit an antiviral response directly in response to a viral infection by releasing type I interferons and stimulating the release of cytokines. In the presence of a minor C/C genotype in the region of the rs1990760 polymorphism of the IFIH1 gene, protein functionality is impaired, leading to a lack of interferon-alpha. A lack of interferon can cause a delay in the primary immune response and an increase in the rate of virus replication and its spread.

The structure of the IFITM3 gene product, which is also a regulator of interferon release, is affected by the genotype in the region of the rs12252 polymorphism. The GG genotype, as a result of alternative splicing, produces a protein truncated by 21 amino acids from the N-terminal region. Such a protein is not functional and, since it is an interferon inducer, leads to an easier entry of the virus into the cell and the risk of a more severe course of the respiratory disease.

The C3AR1 gene is involved in the pathogenesis of complications of respiratory diseases. The C allele (TC or C/C genotype) in the region of rs7842 polymorphism promotes vascular permeability and the release of leukocytes from the vascular bed in lung abscess, which is a complication of pneumonia. This can lead to sepsis.

The TMPRSS2 gene encodes a protein from the family of transmembrane proteases. The A allele in the rs75603675 polymorphism region is a risk factor for severe COVID-19, since the TMPRSS2 protein is associated with facilitating the passage of the SARS-CoV-2 virus.

Transcription factors

The STAT3 gene encodes a protein that is a transcription activator from the STAT family and acts as a regulator of the expression of a number of genes in response to cytokines and growth factors. In addition, STAT3 is involved in the regulation of the body's response to viral and bacterial infections, since the interaction of interleukin-6 with the corresponding receptors triggers the process of STAT3 phosphorylation. The level of interleukin-6 is associated with the severity of the infectious disease and the risk of complications. The G/G genotype in the region of the rs744166 polymorphism in the STAT3 gene plays a protective role. STAT6 is activated inside the cell by foreign nucleic acids, which lead to the activation of innate immunity. Activated in this way, STAT6 regulates a specific set of genes necessary for the recruitment of various immune cells to the site of infection. The minor TT genotype in the region of the rs324011 polymorphism in the STAT6 gene is a risk factor for the severe course of a viral disease.

Receptors

The family of toll-like receptors is an important component of innate immunity, being responsible for the interaction of components of viral particles (proteins or genetic material) with cellular structures. This family includes proteins TLR2, TLR3, TLR4, TLR7, TLR9. They play a fundamental role in pathogen recognition and activation of innate immunity. Activation of TLR-dependent signaling pathways, leading to the secretion of pro-inflammatory cytokines (interleukin-1, interleukin-6, tumor necrosis factor-α, interferon type 1), occurs when various infectious agents enter the body. TLR2/6 and 4 are localized on the cell membrane, while TLR3, TLR7/8 and 9 are localized on the surface of endosomes. The minor GG genotype in the region of the rs1898830 polymorphism in the TLR2 gene has a protective character, since, theoretically, it can enhance the immune response. The minor GG genotype in the rs179008 region in the TLR7 gene is a risk factor for the development of a mild or asymptomatic form of the disease into a more severe one.

Cytokines and chemokines

CCR2 is a key functional receptor for the chemokine CCL2, which in turn regulates T cell inflammatory cytokine expression and T cell differentiation and promotes T cell differentiation into T helper 17 (Th17) during inflammation. Allele A in the region of the rs1799864 polymorphism in the CCR2 gene is a risk factor for viral respiratory infections.

Tumor necrosis factor (TNF, TNF) is a pro-inflammatory cytokine secreted by monocytes and macrophages (not only). It is a pyrogen, causes a feverish state, incl. cachexia, directly or through stimulation by IL-1 secretion. A key mediator of cell death and a participant in the cytokine storm. Allele A in the rs1800629 polymorphism region in the TNF gene is a risk factor for viral respiratory infections.

Table 1. Polymorphisms associated with predisposition to severe viral diseases

Gene Polymorphism Association 1.CCR2 rs1799864 HIV1, HCV, SARS-CoV-2, associated with susceptibility 2. IFIH1 rs1990760 Autoimmune diseases, HCV, SARS-CoV-2, associated with susceptibility, 3.TLR2 rs1898830 Bronchiolitis (RSV), cytomegalovirus infection
Duration of use of oxygen supply 4. STAT3 rs744166 COVID-19; Asthma, lung cancer, 5.STAT6 rs324011 COVID-19 is associated with a severe course; 6.C3AR1 rs7842 SARS-CoV-2, sensitivity, circulatory disorders 7.TMPRSS2 rs75603675 SARS-CoV-2, COVID-19 8.TLR7 rs179008 COVID-19 9. TNF rs1800629 Influenza (risk of severe), asthma, tuberculosis, HIV susceptibility, herpetic encephalitis, COVID-19, 10.IFITM3 rs12252 COVID-19, influenza, herpesviruses

Example #1

Analysis of polymorphisms in 30 genes associated with the emergence and development of COVID-19 in order to identify key factors in the pathogenesis of coronavirus infection.

The authors genotyped 319 genomic DNA samples from patients with varying degrees of severity of COVID-19 disease and 78 control DNA samples from people who had regular or long-term contact with patients with COVID-19, but did not have clinical manifestations and/or antibodies to SARS-CoV-2 . Genotyping was carried out in the area of 34 polymorphisms located in 30 genes. Eight polymorphic markers were identified that were significantly associated with the risk of the disease or its severe course - rs1799864 in the CCR2 gene (OR = 2.21), rs1990760 in the IFIH1 gene (OR = 1.8), rs1800629 in the TNF gene (OR = 1.98), rs75603675 in the gene TMPRSS2 (OR = 1.86), rs7842 in the C3AR1 gene (OR = 2.08), rs179008 in the TLR7 gene (OR = 1.85), rs12252 in the IFIH3 gene (OR = 2.37), and rs324011 in the STAT6 gene (OR = 1.83), as well as two protective against COVID-19 variant - rs744166 in the STAT3 gene (OR = 0.36) and rs1898830 in the TLR2 gene (OR = 0.47).

This example made it possible to predict the risk of a severe course of the respiratory viral disease COVID-19 (10 polymorphisms in 10 genes) and improve the quality of recommendations for the treatment of the disease.

Table 2. Genetic predisposition of a person to a severe course of viral respiratory infections

Gene Polymorphism Interpretation of polymorphism IFIH1 rs1990760 The role of the IFIH1 polymorphism, rs1990760 (C>T) in the epidemiology of viral infection, is well studied, and the minor allele T resists viral infection. Mice with a mutated IFIH1 (946T) protein for this allele have increased interferon production and increased protection against lethal viral infection. Based on mouse and cell culture studies, it is expected that humans carrying the T allele should have higher levels of interferon and a lower risk of SARS-COV2 infection and should be protected from infection. (PMID: 32737579) IFITM3 rs12252 The IFITM3 gene encodes a transmembrane protein that stimulates the production of interferon. The G allele in the rs12252 region is associated with a functionally defective truncated isoform of the protein. A/G and A/A. Similar results were obtained for the Korean population (PMID: 23361009; PMID: 33169083) TNF rs1800629 A powerful pyrogen that induces fever directly or by stimulating interleukin-1 secretion and is involved in the induction of cachexia, under certain conditions it can stimulate cell proliferation and induce cell differentiation. Diseases associated with mutations in the TNF gene include asthma and malaria. Regarding COVID-19: TNF is a pro-inflammatory cytokine that is directly involved in excessive inflammation. An early and high rise in blood TNF levels predicts a high risk of mortality. In this regard, tumor necrosis factor (TNF) inhibition is one of the immunomodulatory approaches that has great promise for the treatment of COVID-19 (PMID: 33294881) TMPRSS2 rs75603675 The serine protease TMPRSS2 is involved in the entry of the SARS-CoV-2 virus into the host cell. Option A in rs35074065 causes overexpression of the protease, which facilitates the entry of the virus into the cell and is a risk factor for increased susceptibility to the SARS-CoV-2 virus. Also, patients who carry the A allele may be potential candidates for treatment with serine protease inhibitors (PMID: 32410502) C3AR1 rs7842 It has been shown that the rs7842 allele in C3AR1 is associated with an increased level of C3AR1 expression and, accordingly, with increased production of C3a is one of the most powerful pro-inflammatory anaphylatoxins produced during complement pathway activation and is involved in the activation of leukocytes and the recruitment of leukocytes to areas of inflammation in the vascular wall (PMID: 25249547). STAT3 rs744166 Allele A is associated with hyperactivation of the JAK-STAT signaling pathway. Since the SARS-Cov-2 virus itself also contributes to the activation of this signaling pathway, the activating genotype also has an additional inhibitory effect on the production of interferon-1. This is associated with the risk of developing complications of the course of COVID-19, such as coagulopathy / thrombosis, pro-inflammatory conditions, profibrotic status and T-cell lymphopenia (PMID: 33037393) STAT6 rs324011 According to the literature data, the minor allele T is associated with an increased level of IgE. This is associated with the risk of various atopic conditions, including bronchial asthma. In addition, the T allele is associated with an increase in the STAT6 gene promoter with the creation of a new nuclear factor-κB transcription factor binding site (PMID: 19665768) TLR7 rs179008 Carriers of the minor TT genotype in rs179008 of the TLR7 gene have significantly lower expression of the interleukin-29 (IL-29) gene, also called interferon-lambda (IFN-λ). This is a risk factor for better susceptibility and more severe viral infections (PMID: 20872712) CCR2 rs1799864 The rs1799864 polymorphism has been well studied in connection with susceptibility to HIV1 infection. It has been shown that in carriers of the G/A and A/A genotypes infected with the HIV1 virus, the progression of infection to AIDS is significantly slower than in carriers of the wild type G/G genotype. So, in the first 4 years after infection with the HIV1 virus, their probability of developing AIDS is reduced by 58%, and in the next 4 years - by 19% compared with carriers of the wild type G/G genotype (PMID 12556692) TLR2 rs1898830 The GG genetic variant of the rs7656411 polymorphism in the TLR2 gene is associated with a risk of acute viral bronchiolitis in various viral respiratory infections. In the situation of infection with the SARS-Cov-2 virus, the TLR-2 receptor recognizes the S-protein of the SARS-Cov-2 virus, but also recognizes HSV proteins and triggers the NF-kappaB signaling pathway through Myd88 and TRAF6, followed by the production of cytokines (PMID: 29253610; PMID : 2925361)

Based on the analysis presented in tables 1 and 2, a genetic panel was created to determine a person's predisposition to a severe course of respiratory infections, which includes a risk ratio depending on the presence of certain alleles and genotypes in polymorphisms of the IFIH1 , IFITM3 , TNF , TMPRSS2 , C3AR1 , STAT3 genes , STAT6 , TLR7 , CCR2 , TLR2 .

Table 3. Genetic panel for predicting the risks of severe viral respiratory diseases and predisposition to them

Gene Polymorphism and genotype in his area Significant genotype Functional value IFIH1 rs1990760; T/C CC, CT Risk IFITM3 rs12252; A/G GG Risk TNF rs1800629; G/A GA, AA Risk TMPRSS2 rs75603675; C/A CA, AA Risk C3AR1 rs7842; T/C TC, CC Risk STAT3 rs744166; A/G AG, GG Protective STAT6 rs324011; C/T CT, TT Risk TLR7 rs179008; A/T TT Risk CCR2 rs1799864; G/A GA, AA Risk TLR2 rs1898830; A/G GG Protective

Example #2

Confirmation of the relationship identified in example 1 polymorphisms with the risk of viral respiratory diseases.

The authors genotyped 98 medical workers (doctors and nurses of infectious diseases departments and intensive care units) who regularly contact patients with respiratory viral infections. Of these, 52 had a reduced incidence of respiratory diseases (ARVI, influenza) - from 0 to 2 times over 5 years (average 1.21 times). In the remaining 46, the incidence was 3 to 4 times over 5 years (average 3.75 times).

Table 4. The results of the study of the frequency of occurrence of minor alleles of polymorphisms according to example 1 in groups with different frequencies of viral infection

gene polymorphism group with reduced incidence high incidence group the number of subjects with a minor allele the frequency of occurrence of the risk allele (protective allele) among the studied the number of subjects with a minor allele the frequency of occurrence of the risk allele (protective allele) among the studied the frequency of occurrence of the risk allele (protective allele) among the European population (data from 1000 Genomes) IFIH1 rs1990760 27 0.519 29 0.630 0.605 IFITM3 rs12252 one 0.019 2 0.043 0.041 TNF rs1800629 5 0.096 6 0.130 0.134 TMPRSS2 rs75603675 21 0.404 nineteen 0.413 0.405 C3AR1 rs7842 15 0.288 thirteen 0.282 ND CCR2 rs1799864 4 0.077 4 0.087 0.087 STAT3 rs744166 33 0.635 eighteen 0.391 0.414 STAT6 rs324011 12 0.231 sixteen 0.348 0.347 TLR7 rs179008 9 0.173 eight 0.174 0.176 TLR2 rs1898830 eighteen 0.346 15 0.326 0.325

It was revealed that the frequency of occurrence of the minor allele in 5 polymorphisms differs in the group with a reduced incidence. This confirms the association of the genotype of the analyzed polymorphic markers with the risk of respiratory viral diseases.

Example #3

Prediction of the risk of occurrence or severe course of a respiratory viral infection or resistance to a respiratory viral infection based on the analysis of the genotype obtained by various methods in the field of polymorphisms given in example 1.

Table 5. Prediction of the risk of occurrence, severity or resistance to respiratory viral infections based on the analysis of the genotype in the area of polymorphisms from example 1.

Gene, polymorphism and genotype variants Genotype at risk or protective genotype significance Points C3AR1 rs7842 (TT/TC/CC) T/C, C/C Risk -2 CCR2 rs1799864 (GG/GA/AA) G/A, A/A Risk -2 TLR7 rs179008 (AA/AG/GG) G/G Risk -one IFIH1 rs1990760 (TT/CT/CC) S/S Risk -2 IFITM3 rs12252 (AA/AG/GG) G/G Risk -3 TMPRSS2 rs75603675 (CC/CA/AA) C/A, A/A Risk -one TNF rs1800629 (GG/GA/AA) G/A, A/A Risk -one STAT6 rs324011 (CC/CT/TT) T/T Risk -one TLR2 rs1898830 (AA/AG/GG) G/G Protective +1 STAT3 rs744166 (AA/AG/GG) G/G Protective +2

From the whole blood of patient No. 1, genomic DNA was isolated and analyzed in the field of polymorphisms from example 1 by the method of capillary sequencing according to Sanger. The following genotype was obtained:

1) C3AR1 rs7842 genotype T/T - 0 points;

2) CCR2 rs1799864 genotype G/A – -2 points;

3) TLR7 rs179008 genotype A/A - 0 points;

4) IFIH1 rs1990760 genotype C/T - 0 points;

5) IFITM3 rs12252 genotype A/A - 0 points.

6) TMPRSS2 rs75603675 genotype С/A – -1 point.

7) TNFrs1800629 genotype G/G – 0 points;

8) STAT6 rs324011 genotype T/T – -1 point.

9) TLR2 rs1898830 genotype A/G – 0 points

10) STAT3 rs744166 genotype A/A - 0 points.

Total score: -4.

Conclusion:

Patient No. 1 has an increased (relative to the general population) risk for the occurrence or severe course of a respiratory viral infection.

From the whole blood of patient No. 2, genomic DNA was isolated and analyzed for polymorphisms from example 1 by PCR with fluorescent probes with real-time detection. The following genotype was obtained:

1) C3AR1 rs7842 genotype T/T - 0 points;

2) CCR2 rs1799864 genotype G/G – -2 points;

3) TLR7 rs179008 genotype A/A - 0 points;

4) IFIH1 rs1990760 genotype C/T – 0 points;

5) IFITM3 rs12252 genotype A/A - 0 points.

6) TMPRSS2 rs75603675 genotype C/C – -1 point.

7) TNFrs1800629 genotype G/G – 0 points;

8) STAT6 rs324011 genotype C/C – -1 point.

9) TLR2 rs1898830 genotype G/G – +1 point

10) STAT3 rs744166 genotype A/A - 0 points.

Total score: +1.

Conclusion:

Patient No. 2 has an increased resistance to the occurrence of either a severe course of a respiratory viral infection or a tendency to an asymptomatic course of a respiratory viral infection.

The method for predicting the risks of a severe course of respiratory diseases of a viral nature and predisposition to them can be carried out both at the stage of the development of the disease and before the development of the disease.

Conclusions about the solution of the set technical problem.

1) A genetic panel has been developed to predict the risks of severe viral respiratory diseases and predisposition to such diseases based on genetic research methods, including the genotype in the area of polymorphisms in genes: rs1990760 in the IFIH1 gene, rs12252 in the IFITM3 gene, rs1800629 in the TNF gene, rs75603675 in the gene TMPRSS2, rs7842 in the C3AR1 gene, rs744166 in the STAT3 gene, rs324011 in the STAT6 gene, rs179008 in the TLR7 gene, rs1799864 in the CCR2 gene, rs1898830 in the TLR2 gene.

2) The possibility of determining the genotype in the field of polymorphisms RS1990760 in the IFIH1, RS1800625 gene in the IFITM3 gene, RS1800629 in the TNF gene, RS75603675 in the C3AR1, RS744166 gene in the STAT3 gene, RS324011 gene in the STAT6 gene, RS17998864 gene in the TLR7 gene in the CCR2 gene, rs1898830 in the TLR2 gene by various methods, including real-time PCR with fluorescent probes and DNA sequencing.

3) The genotype is shown in the RS1990760 polymorphism area in the IFIH1, RS1800622 gene in the IFITM3 gene, RS1800629 in the TNF gene, RS75603675 in the TMPRSS2 gene, RS744166 gene in the STAT3 gene, RS324011 gene in the STAT6 gene, RS179008 gene in the TLR7, RS1799864 gene CCR2 gene, rs1898830 in the TLR2 gene with a risk of occurrence or severe course of a respiratory viral infection or with resistance to a respiratory viral infection.

4) The method of interpreting the results of genotyping in the field of polymorphisms RS1990760 in the IFIH1, RS12252 gene in the iFitm3 gene, RS1800629 in the TNF gene, RS75603675 in the gene TMPRSS2, RS7842 in the C3AR1 gene, RS744166 gene in the STAT6 gene, RS179008 gene in the TLR7 gene, rs1799864 in the CCR2 gene, rs1898830 in the TLR2 gene based on assigning points to the genotype in the area of each polymorphism, summing these points and concluding on the basis of the final sum - sum of points = "0" - general population risk, sum of points less than "0" - increased risk, sum points greater than "0" - the risk is reduced.

Thus, the task set, namely the creation of a method for predicting the risks of severe respiratory diseases of a viral nature in humans and predisposition to them, as well as the creation of a genetic panel for implementing the method by performing a genetic analysis of several polymorphisms at once, has been solved.

Industrial Applicability

All of the above examples confirm the industrial applicability of this invention.

Claims (3)

1. A method for predicting the risks of severe respiratory diseases of a viral nature, which consists in isolating human DNA, determining the DNA sequence in the region of polymorphisms in the human genome: rs1990760 in the IFIH1 gene, rs12252 in the IFITM3 gene, rs1800629 in the TNF gene, rs75603675 in the TMPRSS2 gene, rs7842 in in the C3AR1 gene, rs744166 in the STAT3 gene, rs324011 in the STAT6 gene, rs179008 in the TLR7 gene, rs1799864 in the CCR2 gene, rs1898830 in the TLR2 gene, risk assessment by assigning points to the genotype in the area of each polymorphism, summing these points, while the sum of points = "0 "- the risk is general population, the sum of points is less than "0" - the risk is increased, the sum of points is more than "0" - the risk is reduced, and: genotype - points gene polymorphism homozygous for the major allele heterozygote homozygous for the minor allele CCR2 rs1799864 GG "0" GA "-2" AA "-2" IFIH1 rs1990760 TT "0" TC "0" CC "-2" IFITM3 rs12252 AA "0" AG "0" GG "-3" TNF rs1800629 GG "0" GA "-1" AA "-1" TMPRSS2 rs75603675 CC "0" CA "-1" AA "-1" C3AR1 rs7842 TT "0" TC "-2" CC "-2" STAT3 rs744166 AA "0" AG "0" GG "+2" STAT6 rs324011 CC "0" CT "0" TT "-1" TLR2 rs1898830 AA "0" AG "0" GG "+1" TLR7 rs179008 AA "0" AG "0" GG "-1" 2. The method according to claim 1, which consists in the determination of the DNA sequence in the region of polymorphisms in the human genome by sequencing or by real-time PCR, or by analysis of the length of restriction fragments.