Method and system for providing information on virus infection and / or virus infectious disease

Abstract

To provide information on target virus infection and / or virus infectious disease.SOLUTION: A method includes performing a determination as to detection and / or staging of target virus infection and / or virus infectious disease by using an index related to a target D-amino acid, and based on a result of the determination, providing information on the target virus infection and / or virus infectious disease.SELECTED DRAWING: Figure 1

Description

[Technical field]

[0001] The present invention relates to a method and system for providing information relating to viral infection and / or viral infection. [Background technology]

[0002] Infection refers to the settlement, invasion, and proliferation of pathogens (pathogenic microorganisms) such as viruses in the cells, tissues, and organs of a host through various infection routes. In many cases, pathogens that invade a host are eliminated from the host without being able to infect the host by the body's defense mechanisms, but a state in which infection is established and some symptoms or signs such as fever appear (develop) is called an infectious disease. Infections caused by viruses such as coronaviruses and influenza viruses and infectious diseases caused by them may pose a threat to the medical and economic health due to pandemics. Genetic tests (PCR method, etc.), immunological tests (antigen detection method, etc.), antibody and antigen tests, etc. have been put to practical use in detecting viral infections and viral infectious diseases. In addition, urinary liver fatty acid binding protein (L-FABP) (Patent Document 1) has been proposed as a biomarker for predicting the risk of aggravation of viral infections.

[0003] In recent years, quantitative research has progressed in identifying trace amounts of D-amino acids and L-amino acids in living organisms, including mammals, due to the advancement of technology for identifying and analyzing chiral amino acids. As a result, the existence and functions of some D-amino acids, which have traditionally been treated as total amino acids (D-amino acids + L-amino acids) or conveniently as L-amino acids due to technical limitations, have become clear. It has been shown that in the mammalian intestine, the D-amino acid metabolism of D-amino acid oxidase (DAO) regulates the intestinal immunity of the host against Vibrio bacteria (Non-Patent Document 1). It has also been reported that the %D values ​​({D-amino acids / (D-amino acids + L-amino acids)} x 100) of D-asparagine, D-serine, D-alanine, and D-proline in the blood of humans infected with human immunodeficiency virus (HIV) and receiving antiretroviral therapy correlate with the age and renal function marker (eGFR) of the subjects, but no changes due to HIV infection were observed (Non-Patent Document 2). [Prior art documents] [Patent documents]

[0004] [Patent Document 1] Patent No. 6933834 [Patent Document 2] Patent No. 6868878 [Patent Document 3] Patent No. 6993654 [Patent Document 4] International Publication No. 2020 / 196436 [Patent Document 5] International Publication No. 2013 / 140785 [Non-patent literature]

[0005] [Non-Patent Document 1] Sasabe J, Miyoshi Y, Rakoff-Nahoum S, Zhang T, Mita M, Davis BM, Hamase K, Waldor MK. Interplay between microbial D-amino acids and host D-amino acid oxidase modifies murine mucosal defense and gut microbiota. Nat Microbiol. 2016 Jul 25;1(10):16125. doi: 10.1038 / nmicrobiol.2016.125.

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Summary of the Invention

[0006] Viral infections and the resulting viral infectious diseases have the potential to cause social problems in terms of medical care, economy, and security, as seen in pandemics and bioterrorism, and there is a demand for methods to provide accurate clinical information necessary to deal with these problems. [Means for solving the problem]

[0007] The present inventors comprehensively and highly precisely quantified and analyzed chiral amino acids (amino acids that can be distinguished as D-amino acids and L-amino acids) in the blood of subjects infected with a virus, and found that there is a correlation between the presence or absence of viral infection, the pathology, disease severity, and disease stage of a viral infection, and the amount of chiral amino acids in the blood, and that the fluctuation in the amount of D-amino acids in the blood has a certain pattern. Furthermore, as a result of intensive research into such correlation, they developed an index for D-amino acids in blood as an index for providing information on viral infection and / or viral infection, and found clinical usefulness in clinical testing, diagnosis, and treatment of infectious diseases, leading to the completion of the present invention, which provides a solution to the above-mentioned problems.

[0008] That is, the gist of the present invention relates, for example, to the following: [Item 1] A method for providing information on a subject's viral infection and / or viral infectious disease, comprising: determining whether or not the subject has a viral infection and / or a viral infectious disease using the D-amino acid indicator of the subject; and providing information regarding the subject's viral infection and / or viral disease based on the results of said determination. The method includes: [Item 2] The method according to Item 1, wherein the indicator for D-amino acids is a measured value for D-amino acids in blood or a corrected value or correction formula thereof. [Item 3] The method according to Item 2, wherein the index for the D-amino acid is a value or formula obtained by correcting the amount of the D-amino acid with an index for a substance in the subject's body (e.g., an L-amino acid). [Item 4] The method according to Item 2, wherein the index for the D-amino acid is a value or formula obtained by correcting the amount of the D-amino acid by an index for the renal function of the subject. [Item 5] The method according to any one of Items 1 to 4, wherein the D-amino acid is one or more D-amino acids selected from the group consisting of D-proline, D-serine, D-alanine, and D-asparagine. [Item 6] The method according to any one of Items 1 to 5, wherein the virus is a virus belonging to a family selected from orthomyxovirus, coronavirus, paramyxovirus, rhabdovirus, arenavirus, bunyavirus, filovirus, retrovirus, togavirus, flavivirus, picornavirus, astrovirus, calicivirus, reovirus, parvovirus, adenovirus, papillomavirus, polyomavirus, herpesvirus, hepadnavirus, and poxvirus. [Item 7] The determination of the detection of the viral infection and / or viral infectious disease is determining that the subject is infected with a virus and / or suffers from a viral infection when the indicator for the D-amino acid of the subject shows a downward trend; The method according to any one of items 1 to 6, comprising: [Item 8] The determination of the viral infection and / or stage classification of the viral infection, determining that the subject is in a worsening state of viral infection when the indicator for the D-amino acid of the subject is decreased; determining that the subject is in an improved state of viral infection when the index related to the D-amino acid of the subject is increased; and / or determining that the subject is cured of the viral infection when the index for the D-amino acid of the subject repeatedly decreases and increases and then converges to a healthy reference range; The method according to any one of items 1 to 7, wherein the compound is selected from the group consisting of: [Item 9] The method according to any one of items 1 to 8, wherein the detection and / or stage classification of the viral infection and / or viral infectious disease is carried out by comparing the subject's indicator for the D-amino acid with a determination criterion for viral infection and / or viral infectious disease. [Item 10] Information regarding the subject's viral infection and / or viral disease includes: · detection of viral infection and / or viral infection in said subject; - classifying the stage of viral infection in said subject; Verifying the validity of test results and / or diagnostic results for viral infection and / or viral infection in said subject; and Selecting a treatment for a viral infection in said subject 10. The method according to any one of items 1 to 9, wherein the information is information about an event selected from the group consisting of: [Item 11] The method according to Item 10, wherein the treatment for a viral infection includes a treatment selected from the group consisting of antiviral drugs, blood purification therapy, an artificial respirator, and extracorporeal membrane oxygenation (ECMO). [Item 12] A system for carrying out the method according to any one of items 1 to 11, The system includes an input unit, an analysis and measurement unit, a memory unit, a data processing unit, and an output unit. The input unit inputs information from a subject, the analysis and measurement unit obtains an index for D-amino acids of the subject by analyzing and measuring information from the subject inputted from the input unit; The storage unit stores a determination criterion regarding a virus infection and / or a virus infectious disease, The data processing unit processes the indicators of the subject acquired by the analysis and measurement unit based on the judgment criteria stored in the storage unit, thereby performing a judgment on a viral infection and / or a viral infectious disease of the subject; The output unit outputs a result of the determination by the data processing unit as information regarding a viral infection and / or a viral infection of the subject. The system is configured as follows. Effect of the Invention

[0009] According to the methods and systems of the present invention, by making a judgment using an index related to D-amino acids in a subject, it is possible to provide information regarding a subject's viral infection and / or viral infectious disease with high accuracy and / or efficiency. [Brief description of the drawings]

[0010] [Figure 1] 1 is a graph showing the time course of D-amino acid levels in the blood of influenza virus-infected mice in Example 1. A: D-Ala, B: D-Asn, C: D-Pro, D: D-Ser. [Diagram 2] 2 is a graph showing the amount of D-amino acids in the blood of influenza virus-infected mice in comparison with non-infected mice in Example 1. A: D-Ala, B: D-Asn, C: D-Pro, D: D-Ser. [Diagram 3] 3 is a graph showing the D-amino acid ratio (% D) in the blood of influenza virus-infected mice in comparison with non-infected mice in Example 1. A: D-Ala, B: D-Asn, C: D-Pro, D: D-Ser. [Figure 4] FIG. 4 is a graph showing the body weight of influenza virus-infected mice in Example 1 in comparison with non-infected mice. [Figure 5-1] Figure 5-1 is a graph showing the time course of D-amino acid levels in the blood of coronavirus-infected humans in Example 2. A: D-Ala, B: D-Asn. [Figure 5-2] Figure 5-2 is a graph showing the time course of D-amino acid levels in the blood of coronavirus-infected humans in Example 2. C: D-Pro, D: D-Ser. [Figure 6] 6 is a graph showing the amount of D-amino acids in the blood of coronavirus-infected humans in Example 2 in comparison with non-infected humans. A: D-Ala, B: D-Asn, C: D-Pro, D: D-Ser. [Figure 7] 7 is a graph showing the D-amino acid ratio (%D) in blood of coronavirus-infected humans in Example 2 compared with non-infected humans. A: D-Ala, B: D-Asn, C: D-Pro, D: D-Ser. [Figure 8] Figure 8 is a graph showing the change over time in blood D-amino acid levels in patients with end-stage renal failure among coronavirus-infected humans in Example 2. [Figure 9] 9 shows receiver operating characteristic (ROC) curves of diagnostic ability by univariate analysis of D-amino acid levels in blood of coronavirus-infected humans in Example 2. A: D-Ala, B: D-Pro, C: D-Ser. [Figure 10] 10 is a diagnostic performance ROC curve by univariate analysis of the D-amino acid ratio (%D) in the blood of coronavirus-infected humans in Example 2. A: D-Ala, B: D-Pro, C: D-Ser. [Figure 11] FIG. 11 is a table showing the AUC (area under the curve) and its 95% confidence interval of the diagnostic ability ROC curve obtained by bivariate analysis of D-amino acid levels in the blood of coronavirus-infected humans in Example 2. [Figure 12] FIG. 12 is a block diagram showing a schematic example of a configuration of a system according to the present invention. [Figure 13] FIG. 13 is a flow chart that illustrates an example of processing by the system of the present invention (the method of the present invention). DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0011] The present invention will be described in detail below with reference to specific embodiments. However, the present invention is not limited to the following embodiments, and can be embodied in any form without departing from the spirit of the present invention.

[0012] All patent documents (such as published patent applications and patent publications) and non-patent documents cited in this specification are hereby incorporated by reference in their entirety into this specification.

[0013] In the present specification, amino acids and their residues may be represented by three-letter abbreviations well known to those skilled in the art. The main abbreviations used in the present specification are shown in the following table. [Table 1]

[0014] [Method of Providing Information on Viral Infection and / or Viral Infection (Method of the Invention)] One aspect of the present invention relates to a method for providing information regarding a viral infection and / or viral infection in a subject, the method comprising: using an indicator for a D-amino acid in the subject to determine the detection and / or staging of a viral infection and / or viral infection in the subject; and providing information regarding the viral infection and / or viral infection in the subject based on the results of the determination (hereinafter appropriately abbreviated as the "method of the present invention").

[0015] The method of the present invention is a new evaluation approach for viral infection and / or viral infectious disease, and provides information about the subject's viral infection and / or viral infectious disease by using an index related to D-amino acids (e.g., blood D-amino acid levels) in the subject. This makes it possible to improve the accuracy of detection and / or stage classification of the subject's viral infection and / or viral infectious disease, for example. In addition, the method is a method for assisting in the selection of an appropriate treatment method, for example.

[0016] ·D-amino acids ·L-amino acids In this specification, "D-amino acids" (abbreviated as "D form" where appropriate) and "L-amino acids" (abbreviated as "L form" where appropriate) refer to stereoisomers of amino acids based on the D / L notation system of IUPAC nomenclature. D and L forms are enantiomers. It is known that the majority of proteinogenic amino acids in living organisms are L forms. Although glycine does not have D and L isomers, for the sake of convenience, glycine will be treated as the D form in this specification unless otherwise specified.

[0017] In the present specification, specific examples of D-amino acids include, but are not limited to, glycine, D-alanine, D-histidine, D-isoleucine, D-allo-isoleucine, D-leucine, D-lysine, D-methionine, D-phenylalanine, D-threonine, D-allo-threonine, D-tryptophan, D-valine, D-arginine, D-cysteine, D-glutamine, D-proline, D-tyrosine, D-aspartic acid, D-asparagine, D-glutamic acid, and D-serine. Among them, D-proline, D-serine, D-alanine, and D-asparagine are preferred. These D-amino acids may be used alone or in any combination of two or more.

[0018] In addition, D-cysteine ​​contained in a biological sample is oxidized outside the body and converted to D-cystine, so by measuring an indicator (e.g., amount) of D-cystine instead of D-cysteine, an indicator (e.g., amount) of D-cysteine ​​contained in a biological sample can be calculated.

[0019] ·D-amino acid indicators As used herein, "D-amino acid indicator" refers to any indicator obtained from a living body and having some relationship to D-amino acids (measurement values ​​or test values ​​of D-amino acid indicators obtained from a subject may be simply abbreviated as "D-amino acid test values" or the like.) Examples of D-amino acid indicators include measurement values ​​of D-amino acids in blood or their corrected values ​​or correction formulas.

[0020] An example of a measured value of an index related to D-amino acids in blood is the amount of D-amino acids in blood. In this specification, "amount of D-amino acids in blood" means the amount of D-amino acids contained in a specific amount of blood. The amount of D-amino acids in blood may be expressed as a concentration. The amount of D-amino acids in blood is measured as the amount in a sample of collected blood that has been centrifuged, sedimented, or pretreated for analysis. Therefore, the amount of D-amino acids in blood can be measured as the amount of D-amino acids in a blood sample derived from collected blood, such as whole blood, serum, or plasma. As an example, in the case of analysis using HPLC, the amount of D-amino acids contained in a specified amount of blood is represented by a chromatogram, and can be quantified by comparison with a standard product for peak height, area, and shape, or by analysis using calibration.

[0021] Corrected values ​​or correction formulas for measured values ​​relating to D-amino acids in blood include any value or formula obtained by correcting the amount of D-amino acids in blood. Specific examples include the corrected D / L ratio of the amount of D-amino acids in blood, %D ({D-amino acids / (D-amino acids+L-amino acids)}×100) representing the ratio of D-amino acids in blood, D-amino acid clearance, D-amino acid excretion rate (Non-Patent Documents 3 and 4), and further formulas or values ​​corrected according to the purpose using the amount of D-amino acids as an explanatory variable, and values ​​calculated by a preset formula, etc.

[0022] Another example of the corrected value of the blood D-amino acid amount is a value obtained by adjusting the blood D-amino acid amount according to physiological variables such as age, sex, and BMI. In addition, when the dynamics of D-amino acids are affected by the function of the kidney, a value obtained by adjusting the blood D-amino acid amount according to an index of renal function may be used as the corrected value of the blood D-amino acid amount. Such an index of renal function may be selected from, but is not limited to, creatinine, cystatin C, inulin clearance, creatinine clearance, urinary protein, urinary albumin, β2-MG, α1-MG, NAG, L-FABP, NGAL, glomerular filtration rate, estimated glomerular filtration rate (eGFR), renal function measurements and estimation formulas using D-amino acids (Patent Documents 2, 3, and 4). A specific example is a correction to obtain the ratio of the blood D-amino acid amount to the blood creatinine amount (blood D-amino acid amount / blood creatinine amount). Furthermore, since it is known that D-amino acids in the body fluctuate due to neurodegenerative diseases (such as ALS), autoimmune diseases (such as multiple sclerosis), metabolic diseases (such as diabetes) and the like (Patent Document 5, Non-Patent Document 5), it is also possible to correct the amount of D-amino acids in the blood using the fluctuating factors or markers of each disease.

[0023] Another example of an index related to D-amino acids may be a value or formula obtained by correcting the amount of D-amino acids in the blood of a subject with an index related to a substance in the subject's body (e.g., an L-amino acid). The substance in the subject's body used for correction is not limited to, but may be, for example, the amount of L-amino acids in the blood of the subject or the total amount of amino acids.

[0024] When a value obtained by correcting the amount of D-amino acids in blood by other test values ​​is used as an index for D-amino acids, the criteria and the methods of judgment and analysis for detection and / or stage classification of viral infection and / or viral infectious disease based on the fluctuation (e.g., decrease or increase) of the amount of D-amino acids in blood can be appropriately set or changed according to the correction content for the amount of D-amino acids in blood. As an example, when a value obtained by correcting the amount of D-amino acids in blood as a reciprocal (e.g., [1 / amount of D-amino acids in blood]) or a multiplier is used as an index for D-amino acids, the criteria set based on the fluctuation (e.g., decrease or increase) of the amount of D-amino acids in blood can be used as a reciprocal (e.g., a decrease in the amount of D-amino acids in blood indicates an increase in the value, and an increase in the amount of D-amino acids indicates a decrease in the value) or logarithm.

[0025] In the method of the present invention, any one of D-amino acid indicators (e.g., blood D-amino acid amount, corrected D / L ratio, %D ({D-amino acid / (D-amino acid+L-amino acid)}×100), D-amino acid clearance, D-amino acid excretion rate, etc.) may be used alone or any two or more of them may be used in combination. In the latter case, a panel test in which multiple D-amino acid indicators are combined simultaneously may be used.

[0026] In the method of the present invention, the sample for measuring the indicators related to the D-amino acid of the subject may be one or more samples obtained in a single test, or may be two or more samples obtained in multiple tests. When multiple samples are used, the samples may be obtained at the same time point, or may be obtained at multiple different time points. The form of these samples may be appropriately selected according to various aspects described below.

[0027] The amount of D-amino acids and / or L-amino acids in a sample such as blood can be measured by any method, for example, chiral column chromatography, enzyme methods, or immunological methods using monoclonal antibodies that distinguish optical isomers of amino acids. The amount of D-amino acids and / or L-amino acids in a sample can be measured by any method known to those skilled in the art. Examples include the following chromatographic and enzymatic methods (Y. Nagata et al., Clinical Science, 73 (1987), 105. Analytical Biochemistry, 150 (1985), 238., A. D'Aniello et al., Comparative Biochemistry and Physiology Part B, 66 (1980), 319. Journal of Neurochemistry, 29 (1977), 1053., A. Berneman et al., Journal of Microbial & Biochemical Technology, 2 (2010), 139., WG Gutheil et al., Analytical Biochemistry, 287 (2000), 196., G. Molla et al., Methods in Molecular Biology, 794 (2012), 273., T. Ito et al., Analytical Biochemistry, 371 (2007), 167, etc.), antibody method (T. Ohgusu et al., Analytical Biochemistry, 357 (2006), 15, etc.), gas chromatography (GC) (H. Hasegawa et al., Journal of Mass Spectrometry, 46 (2011), 502., MC Waldhier et al., Analytical and Bioanalytical Chemistry, 394 (2009), 695., A. Hashimoto, T. Nishikawa et al., FEBS Letters, 296 (1992), 33., H.Bruckner and A. Schieber, Biomedical Chromatography, 15 (2001), 166., M. Junge et al., Chirality, 19 (2007), 228., M. C. Waldhier et al., Journal of Chromatography A, 1218 (2011), 4537, etc.), capillary electrophoresis (CE) (H. Miao et al., Analytical Chemistry, 77 (2005), 7190., D. L. Kirschner et al., Analytical Chemistry, 79 (2007), 736., F. Kitagawa, K. Otsuka, Journal of Chromatography B, 879 (2011), 3078., G. Thorsen and J. Bergquist, Journal of Chromatography B, 745 (2000), 389, etc.), high performance liquid chromatography (HPLC) (N. Nimura and T. Kinoshita, Journal of Chromatography, 352 (1986), 169., A. Hashimoto et al., Journal of Chromatography, 582 (1992), 41., H. Bruckner et al., Journal of Chromatography A, 666 (1994), 259., N. Nimura et al., Analytical Biochemistry, 315 (2003), 262., C. Muller et al., Journal of Chromatography A, 1324 (2014), 109., S. Einarsson et al., Analytical Chemistry, 59 (1987), 1191., E. Okuma and H. Abe, Journal of Chromatography B, 660 (1994), 243., Y. Gogami et al., Journal of Chromatography B, 879 (2011), 3259., Y. Nagata et al., Journal of Chromatography, 575 (1992), 147., S. A. Fuchs et al., Clinical Chemistry, 54 (2008), 1443., D. Gordes et al., Amino Acids, 40 (2011), 553., D. Jin et al., Analytical Biochemistry, 269 (1999), 124., J. Z. Min et al., Journal of Chromatography B, 879 (2011), 3220., T. Sakamoto et al., Analytical and Bioanalytical Chemistry, 408 (2016), 517., W. F. Visser et al., Journal of Chromatography A, 1218 (2011), 7130., Y. Xing et al., Analytical and Bioanalytical Chemistry, 408 (2016), 141., K. Imai et al., Biomedical Chromatography, 9 (1995), 106., T. Fukushima et al., Biomedical Chromatography, 9 (1995), 10., R. J. Reischl et al., Journal of Chromatography A, 1218 (2011), 8379., R. J. Reischl and W. Lindner, Journal of Chromatography A, 1269 (2012), 262., S. Karakawa et al., Journal of Pharmaceutical and Biomedical Analysis, 115 (2015), 123., Hamase K, et al., Chromatography 39 (2018) 147-152 etc.) can be mentioned.

[0028] The optical isomer separation and analysis system of the present invention may combine a plurality of separation and analysis. As a specific example, the method for analyzing optical isomers includes the steps of passing a sample containing components having optical isomers through a first column packing material as a stationary phase together with a first liquid as a mobile phase to separate the components of the sample, individually retaining each of the components of the sample in a multi-loop unit, supplying each of the components of the sample individually retained in the multi-loop unit through a flow path to a second column packing material having an optically active center as a stationary phase together with a second liquid as a mobile phase to separate the optical isomers contained in each of the components of the sample, and detecting the optical isomers contained in each of the components of the sample, thereby measuring the amount of D-amino acids and / or L-amino acids in a sample (Japanese Patent No. 4291628). In HPLC analysis, D- and L-amino acids may be derivatized in advance with a fluorescent reagent such as o-phthalaldehyde (OPA) or 4-fluoro-7-nitro-2,1,3-benzoxadiazole (NBD-F), or diastereomerized using N-tert-butyloxycarbonyl-L-cysteine ​​(Boc-L-Cys) or the like (Kenji Hamase and Kiyoshi Zaitsu, Analytical Chemistry, Vol. 53, pp. 677-690 (2004)). Alternatively, the amount of D-amino acids and / or L-amino acids in a sample can be measured by an immunological method using a monoclonal antibody that distinguishes optical isomers of amino acids, for example, a monoclonal antibody that specifically binds to D-amino acids or L-amino acids. In addition, when the total amount of D-amino acids and L-amino acids is used as an index, it is not necessary to separate and analyze D-amino acids and L-amino acids, and amino acids can be analyzed without distinguishing between D-amino acids and L-amino acids. In that case, separation and quantification can be performed by an enzyme method, an antibody method, GC, CE, or HPLC.

[0029] In this specification, the amounts of biomolecules such as D-amino acids, L-amino acids, creatinine, proteins, and drugs are expressed not only in terms of simple mass, weight, or amount of substance (mol), but also in any physical quantity that can be measured, such as the mass, weight, or amount of substance (mol) per tissue, cell, organ, or molecular unit, volume, or weight, or the mass, weight, or amount of substance (mol) in a liquid such as blood or urine, concentration, specific gravity, or density.

[0030] Viruses and viral infections As used herein, the term "virus" refers to a minute infectious structure whose minimum components are nucleic acid carrying genetic information and a protein shell that covers the nucleic acid, and which replicates itself using the cells of other organisms. The viruses to which the present invention can be applied are not particularly limited as long as they infect humans, and may be any virus, including viruses that cause infectious diseases in host organisms through infection.

[0031] Examples of viruses include, but are not limited to, orthomyxoviruses (influenza A virus, influenza B virus, influenza C virus, etc.), coronaviruses (SARS coronavirus (SARS-CoV), MERS coronavirus (MERS-CoV, SARS coronavirus 2 (SARS-CoV-2)), paramyxoviruses (measles virus, mumps virus, etc.), rhabdoviruses, arenaviruses, bunyaviruses, filoviruses, retroviruses (human immunodeficiency virus (HIV)), etc.), togaviruses, flaviviruses (hepatitis C virus, Examples of viruses that can be identified include viruses belonging to various families, such as human adenoviruses (HCV), picornaviruses (rhinovirus types A to C, etc.), astroviruses, caliciviruses, reoviruses, parvoviruses, adenoviruses (human adenovirus types A to G, etc.), papillomaviruses, polyomaviruses, herpesviruses (herpes simplex virus type 1 (HSV-1), herpes simplex virus type 2 (HSV-2), varicella-zoster virus (VZV), etc.), hepadnaviruses (hepatitis B virus (HBV), etc.), and poxviruses (variola virus, cowpox virus, monkeypox virus, camelpox virus, etc.).

[0032] As used herein, "infection" refers to a state in which a pathogen such as a virus adheres to, settles, invades, or grows in the host's cells, tissues, or organs through various infection routes, and includes overt infection, asymptomatic infection, and persistent infection (latent infection). As used herein, "infectious disease" refers to a state in which infection is established and some symptoms or signs appear (for example, fever, chills, headache, muscle pain, joint pain, etc. for influenza, and pneumonia, etc. for COVID-19).

[0033] Examples of viral infections include, but are not limited to, the common cold, influenza, Severe Acute Respiratory Syndrome (SARS), Middle East Respiratory Syndrome (MERS), COVID-19, mumps, other viral pneumonias, chickenpox, shingles, measles, herpes labialis, genital herpes, hepatitis B, hepatitis C, smallpox, and monkeypox.

[0034] ·subject In the present specification, the term "subject" is not limited to, but includes, for example, vertebrates. Examples of vertebrates include mammals, birds, reptiles, amphibians, and fish. Examples of mammals include humans, as well as non-human mammals such as mice, rats, guinea pigs, monkeys, rabbits, cows, horses, pigs, sheep, goats, camels, dogs, and cats. Examples of birds include chickens. Among them, humans or non-human mammals are preferred as subjects, and humans are particularly preferred. In addition, various animals in which virus infection and / or virus infection are induced by transplantation of virus-infected and / or virus-infected cells, genetic modification, or drugs (DNA, RNA, various vaccines, etc.) may be used as subjects. Furthermore, individuals, cells, tissues, organoids, etc. of various animals that serve as a specific virus infection model may be used as subjects.

[0035] ·Judgment criteria According to one embodiment, the method of the present invention compares an index relating to a D-amino acid of a subject with a predetermined criterion to determine whether a subject is infected with a virus and / or has a viral infection. As used herein, the term "criterion" refers to any criterion for determining whether a subject is infected with a virus and / or has a viral infection using an index relating to a D-amino acid of a subject. Examples include, but are not limited to, a predetermined reference value consisting of a single numerical value (suitably referred to as a "criterion value") and a predetermined reference range defined by an upper limit and a lower limit (suitably referred to as a "criterion range"). That is, as used herein, the term "criterion" encompasses both "criterion value" and "criterion range".

[0036] The method of comparing the target D-amino acid index with a predetermined criterion is not particularly limited. When a criterion value is used as the criterion, for example, the criterion value is used as the upper limit of the D-amino acid index, and the criterion can be whether the target D-amino acid index is equal to or greater than the criterion value, or whether it exceeds the criterion value. Alternatively, the criterion value is used as the lower limit of the D-amino acid index, and the criterion can be whether the target D-amino acid index is equal to or less than the criterion value, or whether it falls below the criterion value. On the other hand, when a criterion range is used as the criterion, for example, the criterion can be whether the target D-amino acid index is within the criterion range, exceeds the criterion range, or falls below the criterion range. Alternatively, the judgment can be made based on whether the indicator for the target D-amino acid fluctuates over time from outside the judgment criteria range to within the judgment criteria range, fluctuates from within the judgment criteria range to outside the judgment criteria range, remains outside the judgment criteria range, or remains within the judgment criteria range.

[0037] In the method of the present invention, a single judgment reference value or judgment reference range may be used as the judgment criterion, or multiple judgment reference values ​​or judgment reference ranges may be used in combination, or one or more judgment reference values ​​and one or more judgment reference ranges may be used in combination.

[0038] Detection of viral infection and / or viral disease According to one aspect, the method of the present invention comprises using an indicator for a subject's D-amino acid to determine whether a subject's viral infection and / or viral infection has been detected, and providing information relating to the detection of a viral infection and / or viral infection in the subject based on the results of the determination.

[0039] Specifically, a subject's viral infection and / or viral infection can be detected by the change in the indicator of D-amino acid in the subject, and information on the detection result can be provided. For example, in a subject suspected of influenza infection (e.g., a patient, a medical worker, etc.), when the test value of the blood D-amino acid amount, which is one of the indicators of D-amino acid in the subject, decreases, the subject can be judged to be infected with a virus and / or suffering from a viral infection, and the judgment result can be provided as detection information of viral infection and / or viral infection. In addition, in a subject suspected of influenza infection, when the test value of the blood D-amino acid amount shows a tendency to decrease, the subject can be judged to be infected with a virus and / or suffering from a viral infection, and the judgment result can be provided as detection information of viral infection and / or viral infection.

[0040] In this case, the above detection may be performed by comparing the test value of the subject's D-amino acid indicator with a criterion determined from the D-amino acid indicator in the past healthy state of the same subject, or with a criterion determined from the D-amino acid indicator of a patient or patient group known to have a viral infection and / or a viral infection. For example, the presence or absence of detection of a viral infection can be determined by comparing the test value of the subject's D-amino acid indicator with a preset criterion (such as a reference range or clinical judgment value) for the D-amino acid indicator, utilizing the fact that the profile of D-amino acids and / or L-amino acids in the blood of a subject with a viral infection and / or a viral infection is different from the profile of D-amino acids and / or L-amino acids in the blood of a subject without a viral infection and / or a viral infection. As a specific example, the amount of D-amino acids in blood is used as the D-amino acid indicator, and when the test value of the amount of D-amino acids in the blood of the subject is lower than a predetermined criterion or shows a tendency to decrease more than the predetermined criterion, the subject can be determined to be positive for a viral infection and / or a viral infection.

[0041] When a value obtained by correcting the amount of D-amino acids in blood using other test values ​​or the like is used as an index for D-amino acids, the criteria for detecting viral infection and / or viral infection based on the fluctuation (e.g., decrease or increase) of the amount of D-amino acids in blood and the methods for judgment and analysis can be appropriately set or changed according to the correction content for the amount of D-amino acids in blood. As an example, when a value obtained by correcting the amount of D-amino acids in blood using a reciprocal or multiplier is used as an index for D-amino acids, the criteria set based on the fluctuation (e.g., decrease or increase) of the amount of D-amino acids in blood can be used as a reciprocal or logarithm.

[0042] There have been no reports to date of changes in D-amino acid indicators (e.g., a decrease or a tendency toward a decrease in blood D-amino acid levels) in subjects with viral infection and / or viral infection, and it has not been known that there is a relationship between the presence or absence of viral infection and / or viral infection and changes in D-amino acid indicators (e.g., a decrease or a tendency toward a decrease in blood D-amino acid levels). Thus, the method of the present invention is extremely useful in clinical detection and diagnosis of viral infection and / or viral infection.

[0043] Viral infection and / or staging of viral infection According to one aspect, the method of the present invention comprises using an index related to a D-amino acid in a subject to determine the stage of a viral infection and / or a viral infectious disease in the subject, and providing information relating to the stage of a viral infection and / or a viral infectious disease in the subject based on the results of the determination.

[0044] As used herein, the worsening of symptoms of a viral infection is referred to as "worsening," and depending on the degree of worsening, it is referred to as "moderate" or "severe" (for example, in the case of COVID-19, a state in which an artificial respirator or ECMO is required). As used herein, the reduction in the severity of symptoms is referred to as "improvement," "recovery," or "remission," and the state in which the pathogen has been eliminated from the body is referred to as "cure." As used herein, the course, results, and outlook of symptoms are referred to as "outcome."

[0045] In this specification, "disease stage classification" of a viral infection refers to classifying the disease stage of a target infection into "latent stage," "acute stage (worsening stage)," "recovery stage (improvement stage)," "cured," "aftereffect stage," etc., according to the degree of progression. Generally, classification is based on genetic testing (molecular biological testing), immunological testing, diagnostic imaging, and physical findings, and serves as the basis for determining severity, treatment methods, and prognosis evaluation. The definitions and standards of disease stages may be amended or standardized based on the handling rules for various infectious diseases established by related academic organizations and societies.

[0046] In such an embodiment, the method of the present invention comprises: determining that the subject is in a worsening state of viral infection when the indicator for the D-amino acid of the subject is decreased; determining that the subject is in an improved state of viral infection when the index related to the D-amino acid of the subject is increased; and / or determining that the subject is cured of the viral infection when the index for the D-amino acid of the subject repeatedly decreases and increases and then converges to a healthy reference range; may also include

[0047] In addition, in such an embodiment, the stage classification may be performed by comparing the test values ​​of the D-amino acid indicators of the subject with judgment criteria determined from the D-amino acid indicators of viral infection patients whose viral infection stage has been classified.

[0048] Specifically, for example, the stage of a viral infection can be classified using the test value of the blood D-amino acid amount, which is a type of index related to D-amino acids of a subject. That is, by utilizing the fact that the profile of D-amino acids and L-amino acids in the blood of a subject with a viral infection varies depending on the progress of the viral infection, the test value of the index related to D-amino acids of the subject can be compared with a predetermined judgment reference value (reference range or clinical judgment value, etc.) based on the amount of D-amino acids in the blood, and the stage of the viral infection can be determined, and information on the corresponding treatment and outcome can be provided. For example, the index related to D-amino acids in blood can be used to compare the stage of a viral infection with the test value of the subject, for which a judgment reference value has been set, to determine the stage of the viral infection of the subject, and the obtained judgment result can be provided as information on the stage of the viral infection of the subject.

[0049] Also, for example, the stage of a viral infection of a subject can be classified by the change in the test value of the blood D-amino acid amount, which is a kind of index for the D-amino acid of the subject. That is, a period in which the test value of the blood D-amino acid amount of a subject shows a decrease and / or a tendency to decrease can be classified as the subject being in the acute phase of a viral infection, a state or stage showing worsening and / or aggravation, or a state or stage showing no improvement and / or recovery. Furthermore, a period in which the test value of the blood D-amino acid amount of a subject shows an increase and / or a tendency to increase can be classified as the subject being in the recovery phase of a viral infection, a state or stage showing improvement and / or recovery. In one embodiment, when the blood D-amino acid amount of a subject shows a transient decrease, increase, or repetition thereof, and then converges to the reference range for a healthy state, the subject can be classified as being in remission, good outcome, or cured phase of a viral infection. Furthermore, since viral infections progress from mild to moderate to severe as the condition continues to worsen, the severity can also be classified based on the duration of the disease, the degree of fluctuation in the amount of D-amino acids in the blood, etc.

[0050] According to one embodiment, when a test value of the blood D-amino acid amount, which is one of the indicators of D-amino acids, of a subject is maintained at a decreased level, the subject can be judged to be in a state where the viral infection has worsened and / or become severe, or the viral infection has not improved and / or recovered. When a test value of the blood D-amino acid amount of a subject is temporarily decreased and then increases, the subject can be judged to be in a state where the viral infection has improved or has recovered.

[0051] Furthermore, according to one embodiment, when a test value of a subject's D-amino acid indicator shows a transient decrease or increase, or a repetition of these, and then converges to a reference range for the indicator in the same subject's past healthy state, or to a reference range for the indicator set by a specified reference population in a healthy state, the subject can be determined to be in remission, with a good outcome, or cured of the infectious disease.

[0052] A decrease or increase in a subject's test value of an index related to D-amino acids can be determined, for example, by comparing the test value of the subject's index with a reference value or reference range of the index in the same subject's past healthy state, a reference value or reference range of the index set by a reference population in a predetermined healthy state, or a predetermined clinical judgment value, or by comparing test values ​​of the subject at two or more points over time.

[0053] When a value obtained by correcting the amount of D-amino acids in blood using other test values ​​or the like is used as an index for D-amino acids, the criteria for classification of viral infection and / or stage of viral infection based on the fluctuation (e.g., decrease or increase) of the amount of D-amino acids in blood and the methods for judgment and analysis can be appropriately set or changed according to the correction content for the amount of D-amino acids in blood. As an example, when a value obtained by correcting the amount of D-amino acids in blood using a reciprocal or multiplier is used as an index for D-amino acids, the judgment criteria set based on the fluctuation (e.g., decrease or increase) of the amount of D-amino acids in blood can be used as a reciprocal or logarithm.

[0054] There have been no reports to date of any change in D-amino acid indicators (e.g., a pattern of decrease or increase in blood D-amino acid levels) in subjects with viral infection and / or viral infection depending on the stage of the viral infection and / or viral infection, and it has not been known that there is a relationship between the stage of viral infection and / or viral infection and changes in D-amino acid indicators (e.g., a pattern of decrease or increase in blood D-amino acid levels). Thus, the method of the present invention is extremely useful in clinical detection and diagnosis of viral infection and / or viral infection.

[0055] -Verifying the validity of test results and / or diagnostic results for viral infection and / or viral infection The method of the present invention can also provide other information based on the results of the detection and / or staging of the viral infection and / or viral infectious disease. For example, according to one embodiment, the method of the present invention can provide information regarding the verification of the validity of the test results and / or diagnosis results of the viral infection and / or viral infectious disease of the subject based on the results of the detection and / or staging of the viral infection and / or viral infectious disease of the subject, which are performed based on the test values ​​of the indicators related to D-amino acids of the subject.

[0056] PCR tests and antibody tests commonly used for viral infection tests have the problem that the amount of virus or protein contained in the collected sample (nose or throat swabs, sputum, etc.) is often below the measurement limit, resulting in false negatives (Non-Patent Documents 6 and 7). The amount of D-amino acids in blood fluctuates due to the influence of viral infection on its intake, absorption, transport, distribution, metabolism (synthesis and decomposition), excretion, action, etc., and therefore the fluctuation of the test value of the D-amino acid indicator, such as the amount of D-amino acids in the blood of the subject, is fundamentally different from the fluctuation of conventional test markers, and is therefore highly useful in determining true positives and false positives. In other words, if a subject is suspected of viral infection based on the subject's environment, symptoms, etc., but a negative result is obtained in a specified test, the test value of the D-amino acid indicator can be used to determine whether the subject is false negative (type II error) or true negative. In particular, the phenomenon of a decrease (reduction) in the amount of D-amino acids in a subject's blood is specific to viral infection. Therefore, unlike direct detection of viruses or their fragments, indicators for D-amino acids that can distinguish between D-amino acids and L-amino acids in physiological changes in the host and examine the phenotype of a disease state have special characteristics and are effective in detecting viral infections and / or determining the authenticity of test results.

[0057] In this specification, "verification of the validity of test or diagnostic results" refers to the verification of the validity of a subject diagnosed by a clinical test that may include false positive or false negative judgments, such as a medical interview or tests of specimens (pharyngeal secretions, phlegm / airway secretions, urine, vaginal secretions, feces, blood, cerebrospinal fluid, etc.) collected from a subject (biochemical tests, serological tests, endocrine tests, microbiological tests, virological tests, culture tests, microscopic tests, genetic tests (PCR method, hybridization, etc.), immunological tests (antigen / antibody detection methods), pathological tests, imaging tests (endoscopic tests, contrast agent tests, ultrasound tests, CT tests, MRI tests, etc.), and companion diagnostic tests that investigate the effects and side effects of specific drugs in advance, etc., using different principles. It means verifying the validity of a diagnostic result by using an index. As a specific example, for a subject determined to be positive by antibody detection of a predetermined paired serum, if the subject's viral infection and / or viral infectious disease is determined to be positive based on the determination using an index related to D-amino acids such as the amount of D-amino acids in the subject's blood, it can be determined to be a true positive, and if it is determined to be negative, it can be determined to be a false positive (type I error). Conversely, for a subject determined to be negative, if the subject's viral infection and / or viral infectious disease is determined to be positive based on the determination using an index related to D-amino acids such as the amount of D-amino acids in the subject's blood, it can be determined to be a false negative (type II error), and if it is determined to be negative, it can be determined to be a true negative.

[0058] The validity of the test results and / or diagnostic results based on the results of the detection and / or stage classification of the viral infection and / or viral infectious disease is verified by comparison or analysis using the criteria for the indicator (also referred to as "reference range" or "clinical judgment value" in this specification). The criteria (reference range or clinical judgment value) that can be used in the present invention are set as individual reference values ​​and clinical judgment values ​​based on the test values ​​in the healthy state of each subject, or are set as generally the 95% interval in the middle of the test value distribution of healthy individuals (reference individuals) who meet certain criteria or a group of subjects with infection and infectious diseases, but any interval can also be set depending on the purpose. The criteria are generally used to judge the diagnosis, prevention, treatment, and prognosis of a specific pathology, and include diagnostic thresholds, treatment thresholds, and preventive medicine thresholds. These thresholds (cut-off values) can be set by utilizing analytical data or results on predictive ability and diagnostic ability using ROC curves (Receiver Operating Characteristic curves), multivariate logistic regression models, Cox proportional hazard models, etc., and by case-control studies, clinical medical empirical rules, case accumulation studies, cohort studies, expert consensus, etc. For example, by comparing the above-mentioned various test values ​​with the determination results based on the D-amino acid index by the method of the present invention, information on the verification results of the validity of the test or diagnostic results of the subject can be provided. In addition, in the test values ​​of the D-amino acid index of the subject at any two or more points in time, if the test values ​​show an increase over time as an increasing trend, and if the test values ​​show a decrease over time as a decreasing trend, the test values ​​of the D-amino acid index of the subject at those two or more points in time can be compared and analyzed to verify the validity of the test results and / or diagnostic results in more detail.

[0059] Viral infection and / or the choice of treatment for viral infection In addition, according to one embodiment, the method of the present invention can also provide information regarding the selection of a treatment for a viral infection in a subject based on the results of the detection and / or staging of the aforementioned viral infection and / or viral infectious disease, which is performed based on the test values ​​of indicators related to D-amino acids in the subject.

[0060] As used herein, "selection of a therapeutic means" refers to selecting the most suitable means from surgery, radiation therapy, chemotherapy, drug therapy, immunotherapy, dietary therapy, exercise therapy, etc., and further each technique (e.g., surgical procedure, administration method, etc.), or determining the priority thereof, for a subject diagnosed with a specific disease, and also to administering treatment to the subject so that a predetermined therapeutic means can be selected. The criteria and objectives for selection include cure of disease, relief or elimination of symptoms, halting or slowing the progression of disease, prevention of disease or symptoms, suppression of deterioration of underlying disease, avoidance or minimization of side effects, cost-effectiveness, improvement or maintenance of QOL, etc.

[0061] According to one embodiment, prior to carrying out the method of the present invention, subjects known to have a viral infection are clearly distinguished into those who will respond to a treatment for the viral infection and / or those who will not respond, and criteria for selecting a treatment are set in advance based on the measured values ​​of the D-amino acid indicators of these subjects. Then, when carrying out the method of the present invention, the test values ​​of the D-amino acid indicators obtained from a new subject are compared with the criteria set based on the known subjects, and the obtained results can be provided as information regarding the viral infection of the subject and / or the selection of a treatment for the viral infection.

[0062] According to one embodiment, the method of the present invention can provide information to assist in the selection of a treatment for a viral infection using an index related to a subject's D-amino acid. By utilizing the fact that the profiles of D-amino acids and L-amino acids in the subject's blood differ in relation to the response and change of the viral infection during viral infection and treatment, and by comparing the test value of the index related to the D-amino acid of the subject with a preset judgment criterion (reference range or clinical judgment value), the optimal treatment can be selected from drug therapy, oxygen therapy (nasal cannula, HFNC, CPAP, NPPV, etc., respiratory failure), surgical treatment, artificial respiratory management (artificial ventilator, etc.), ECMO (extracorporeal membrane oxygenation), blood purification therapy (dialysis, plasma exchange, apheresis, etc.), thrombosis countermeasures, renal disorder countermeasures, symptomatic treatment (antipyretic, antitussive, etc.), dietary therapy, etc., or the determination of the priority of the treatment can be assisted. By monitoring the index related to the D-amino acid of the subject in real time even during the treatment stage, information to assist in the selection of the next stage of treatment can be provided. Furthermore, in the event of a pandemic or bioterrorism, D-amino acid indicators can be used for triage, which determines and selects the priority of medical care and treatment for the target, and for risk assessment in each pandemic phase of the alert, pandemic, and transitional stages. Antiviral drugs mainly used in drug therapy are drugs that have effects such as inhibiting the adsorption and invasion of viruses into host cells, inhibiting intracellular uncoating, inhibiting nucleic acid synthesis, inhibiting protein synthesis, and inhibiting extracellular release, and oxygen therapy, the use of artificial ventilators, and ECMO are used for severely ill subjects.

[0063] According to one embodiment, the method of the present invention can provide information to select an optimal drug such as an antiviral drug as a means for treating a viral infection, or to assist in determining the priority of the drug, using an index related to the D-amino acid of the subject. As a specific example, the index related to the D-amino acid of the subject can be used to set a criterion (reference range or clinical diagnostic value) for the effect, side effects, and adverse reactions of a given drug in advance, and compare the criterion with the test value of the subject to consider whether or not to administer the drug. According to one embodiment, the index related to the D-amino acid of the subject can be used to predict and determine the effect, side effects, and adverse reactions after drug administration, or to provide information to assist in determining whether or not to continue or stop administration, or the dosage and timing of administration. Furthermore, the index related to the D-amino acid of the subject can be used to provide information to assist in determining a means for adjusting the value of the index related to the D-amino acid in the subject. Furthermore, the index related to the D-amino acid of the subject can be used to provide information for screening a means for adjusting the value of the index related to the D-amino acid in the subject.

[0064] Specific examples of drugs that can be used as a means of treating viral infections include, but are not limited to, the following: Drugs for treating viral infections include neuraminidase inhibitors (oseltamivir, zanamivir, peramivir, laninamivir, etc.), M2 protein inhibitors (amantadine, etc.), RNA polymerase inhibitors (favipiravir, molnupiravir, etc.), cap-dependent endonuclease inhibitors (baloxavir, marboxil, etc.), anti-herpes virus drugs (acyclovir, valacyclovir, famciclovir, amenamevir, etc.), anti-cytomegalovirus drugs (anti-mycobacterial drugs, anti ... Antiviral drugs (ganciclovir, foscarnet, valganciclovir, etc.), anti-hepatitis B virus drugs (entecavir, tenofovir, lamivudine, adefovir, etc.), anti-hepatitis C virus drugs (sofosbuvir, ribavirin, redibasvir, etc.), nucleoside reverse transcriptase inhibitors (tenofovir, emtricitabine, etc.), non-nucleoside reverse transcriptase inhibitors (rilpipridine, efavirenz, etc.), integrase inhibitors (elvitegravir, dolutegravir, vir, etc.), protease inhibitors (darunavir, ritonavir, etc.), CCR5 inhibitors (maraviroc, etc.), antibiotics, herbal medicines (kakkonto, shoseiryuto, maoto, etc.), acetaminophen, NSAIDs, antihistamines, immunosuppressants (steroids, baricitinib, etc.), neutralizing antibody drugs (remdesivir, casirivimab, imdevimab, sotrovimab, etc.), Janus kinase inhibitors (baricitinib), RNA polymerase inhibitors (remdesivir), biologically active peptides Antibodies that may be used include peptides (adrenomedullin, etc.), GM-CSF preparations (sargramostim, etc.), anticoagulants (heparin, etc.), antiparasitic drugs (ivermectin, etc.), humanized anti-human IL-6 receptor monoclonal antibodies (tocilizumab, sarilumab, etc.), serine protease inhibitors (nafamostat, etc.), vaccines (DNA vaccines, RNA vaccines, attenuated vaccines, adenovirus vector vaccines, etc.), oral fluid replacement, infusions, blood transfusions, etc.

[0065] ·others The method of the present invention can be used to provide various other information. For example, according to one embodiment, the method of the present invention can provide information on screening for viral infection and / or viral infection and diagnosis of the pathology based on the results of the detection and / or stage classification of the aforementioned viral infection and / or viral infection, which are performed based on the test values ​​of the indicators related to the D-amino acid of the subject. In addition, according to one embodiment, the method of the present invention can provide information on screening of efficacy, side effects, and adverse reactions in drug development, judgment of clinical trials, alternative endpoints, etc., based on the results of the detection and / or stage classification of the aforementioned viral infection and / or viral infection, which are performed based on the test values ​​of the indicators related to the D-amino acid of the subject. The multiple pieces of information may be provided individually or simultaneously depending on the purpose.

[0066] [System for Providing Information on Viral Infection and / or Viral Infection (System of the Present Invention)] One aspect of the present invention relates to a system for providing information regarding viral infection and / or viral infectious disease in a subject by carrying out the method of the present invention (suitably abbreviated as "the system of the present invention").

[0067] FIG. 12 is a block diagram showing a schematic example of the configuration of the system of the present invention. However, the configuration shown in FIG. 12 is merely an example, and the configuration of the system of the present invention is not limited thereto. The sample analysis system 10 shown in FIG. 12 includes a memory unit 11, an input unit 12, an analysis and measurement unit 13, a data processing unit 14, and an output unit 15. The memory unit 11 is configured to store various information including criteria for determining viral infection and / or viral infection. The input unit 12 is configured to input various information including information from the subject. The analysis and measurement unit 13 is configured to perform various analysis and measurement, such as obtaining an indicator for the D-amino acid of the subject by analyzing and measuring the information from the subject. The data processing unit 14 is configured to perform various calculation processes, such as performing a determination of viral infection and / or viral infection of the subject by processing the indicator for the D-amino acid of the subject based on the criteria. The output unit 15 is configured to output various information including information regarding viral infection and / or viral infection.

[0068] Specifically, the storage unit 11 is configured with, for example, a memory device such as a RAM, a ROM, or a flash memory, a fixed disk device such as a hard disk drive, or a portable storage device such as a flexible disk or an optical disk. The storage unit 11 is configured to store various information such as data and instructions input from the input unit 12, data measured by the analysis and measurement unit 13, the results of arithmetic processing performed by the data processing unit 14, and computer programs and databases used for various processes of the information processing device that realizes the sample analysis system 10. The computer program may be installed from a computer-readable recording medium such as a CD-ROM or a DVD-ROM, or via the Internet. The computer program is installed in the storage unit 11 using a known setup program or the like.

[0069] The input unit 12 is an interface with the outside of the sample analysis system 10, and also includes an operation unit such as a keyboard and a mouse. This allows the input unit 12 to input data measured by the analysis and measurement unit 13, instructions for arithmetic processing to be performed by the data processing unit 14, and the like. Furthermore, when the analysis and measurement unit 13 is located externally, the input unit 12 may include an interface unit that can input the measured data, etc. via a network or a storage medium, separate from the operation unit.

[0070] The analysis and measurement unit 13 is configured to obtain an index related to the D-amino acid of the subject by analyzing and measuring information from the subject. For example, the analysis and measurement unit 13 can be configured to measure at least the amount of D-amino acid from a blood sample of the subject. Therefore, the analysis and measurement unit 13 may have a configuration that enables separation and measurement of D- and L-isomers of amino acids. The amino acids may be configured to analyze one by one, or may be configured to analyze some or all types of amino acids together. The analysis and measurement unit 13 is not intended to be limited to the following, but may be, for example, a chiral chromatography system equipped with a sample introduction unit, an optical resolution column, and a detection unit, preferably a high performance liquid chromatography system. From the viewpoint of detecting only the amount of a specific amino acid, quantification may be performed by an enzyme method or an immunological method. The analysis and measurement unit 13 may be configured separately from the evaluation system of the test value, and the measured data, etc. may be input via the input unit 12 using a network or a storage medium.

[0071] The data processing unit 14 can select information on the viral infection and / or viral infection of the subject by comparing the indicator for D-amino acids measured by the analysis and measurement unit 13 with the judgment criteria stored in the storage unit. The indicator for D-amino acids may be a formula or value corrected by the amount of a substance in the subject's body (e.g., the amount of D-amino acids or an index of a test), or may be a formula or value corrected by physiological fluctuation factors such as age, sex, BMI, etc. The data processing unit 14 executes various arithmetic processing on the data measured by the analysis and measurement unit 13 and stored in the storage unit 11 according to a program stored in the storage unit. The arithmetic processing is performed by a CPU included in the data processing unit. This CPU includes a functional module that controls the analysis and measurement unit 13, the input unit 12, the storage unit 11, and the output unit 15, and can perform various controls. Each of these units may be composed of an independent integrated circuit, microprocessor, software, etc.

[0072] The output unit 15 is configured to output information on the viral infection and / or viral infection of the subject, which is the result of the arithmetic processing performed by the data processing unit. The output unit 15 may be an output means such as a display device such as a liquid crystal display that directly displays the results of the arithmetic processing, or a printer, or may be an interface unit for outputting to an external storage device or via a network.

[0073] FIG. 13 is a flow chart showing an example of the processing by the system of the present invention (the method of the present invention). However, the processing shown in FIG. 13 is merely an example, and the processing by the system of the present invention is not limited thereto. First, the criteria for viral infection and / or viral infection are read from the input unit 12 and stored in the memory unit 11 (step S1). Next, information on the D-amino acid of the subject is read from the input unit 12 and stored in the memory unit 11 (step S2). Then, the analysis and measurement unit 13 analyzes and measures the information from the subject stored in the memory unit 11 to obtain an index for the D-amino acid of the subject (step S3). Next, the data processing unit 14 processes the index for the D-amino acid of the subject obtained by the analysis and measurement unit 13 based on the criteria stored in the memory unit 11 to perform a judgment on the viral infection and / or viral infection of the subject (step S4). Next, the determination result of the subject's viral infection and / or viral infection by the data processing unit 14 is stored in the memory unit 11 as information on the subject's viral infection and / or viral infection, and is output from the output unit 15 (step S5).

[0074] [others] Although the present invention has been described in detail above with reference to specific embodiments, the present invention is not limited to these embodiments. Those skilled in the art can derive various other inventive concepts from the above description, and all of these are included in the technical scope of the present invention.

[0075] For example, it is possible to provide a computer program (appropriately referred to as the "program of the present invention") for implementing the system of the present invention using a general-purpose information processing device. Specifically, the program of the present invention can be configured as a program including computer instructions that can cause the information processing device and external devices such as an input / output interface and an analyzer connected thereto to function as a sample analysis system 10 including a memory unit 11, an input unit 12, an analysis and measurement unit 13, a data processing unit 14, and an output unit 15, for example, as shown in FIG. 12, by installing and executing the program of the present invention on a general-purpose information processing device. Such a program of the present invention can be realized by knowledge of computer programming well known to those skilled in the art. Such a program of the present invention and a recording medium such as a CD-ROM including the program are also included in the technical scope of the present invention.

[0076] Since the present invention can be carried out by comparing the test value of an indicator related to D-amino acids in a subject with a predetermined judgment standard (reference range or clinical judgment value), it can be carried out by persons other than physicians, such as clinical testing, health examination, and data processing companies, or by analysis and analysis systems and analysis and analysis programs, without the need for judgment by a physician, and thus does not fall under so-called medical procedures, etc. In other words, since the present invention provides a judgment result for the detection and / or stage classification of a viral infection and / or viral infection in a subject based on an indicator related to the D-amino acids in the subject, it does not replace medical procedures such as diagnosis and treatment by a physician, but has extremely high technical usefulness as a preliminary or auxiliary method for improving the accuracy and efficiency of such diagnosis and treatment. EXAMPLES

[0077] The present invention will be described in more detail below with reference to examples, but these examples are merely illustrative and are not intended to limit the present invention in any way. Those skilled in the art can easily modify and alter the present invention based on the description in this specification, and all such modifications and alterations are within the technical scope of the present invention.

[0078] In the examples, the symbols have the following meanings: PD-AA: Plasma D-amino acid amount (nmol / mL, μM) PL-AA: L-amino acid concentration in plasma (nmol / mL, μM) PCre: plasma creatinine concentration P%D:(PD-AA / PD-AA+PL-AA)×100(%)

[0079] [Example 1: Influenza model mouse test] To induce severe infection in a short period of time in C57BL6 mice (SLC, Tokyo, Japan) raised in a specific pathogen control facility, anesthetized 4-week-old mice (n=7) were infected intranasally with 10 L of water containing 50 times the TCID50 (median tissue culture infectious dose) of PR8 with the H1N1 influenza virus strain (ATCC, Manassas, USA). After infection, blood was collected from each mouse and control mice (n=5) at a predetermined schedule, and chiral amino acids (D- and L-amino acids) in plasma were quantitatively analyzed by 2D-HPLC.

[0080] This experiment was approved by the Animal Committee of the National Institutes of Biomedical Innovation, Health and Nutrition and was carried out in accordance with the guidelines of the Animal Protection and Management Act of Japan.

[0081] FIG. 1 is a graph showing the time course of blood D-amino acid levels in influenza virus-infected mice in Example 1. FIG. 1A shows data for PD-Ala, FIG. 1B shows data for PD-Asn, FIG. 1C shows data for PD-Pro, and FIG. 1D shows data for PD-Ser. As is clear from these data, the blood D-amino acid levels of influenza virus-infected mice all decreased (reduced) over time after infection, making it possible to provide information on viral infection based on the fluctuation in blood D-amino acid levels. Furthermore, the blood D-amino acid levels of influenza virus-infected mice showed a continuous decrease without increasing, and during that time symptoms (weight loss, see FIG. 4) continued, worsening, and the mice died, so it can be determined that the influenza virus-infected mice were in the acute phase (worsening phase) throughout the test period after infection.

[0082] FIG. 2 is a graph showing a t-test (*<0.05) performed on PD-AA in the influenza virus-infected mouse group and the non-infected mouse (control) group in Example 1. FIG. 2A shows data for PD-Ala, FIG. 2B shows data for PD-Asn, FIG. 2C shows data for PD-Pro, and FIG. 2D shows data for PD-Ser. These data show that PD-AA in influenza virus-infected mice is significantly decreased (reduced) compared to the control group, indicating that viral infection can be determined based on a decrease in the amount of D-amino acids in the blood. A more accurate determination can be achieved by performing a panel test using multiple PD-AA.

[0083] FIG. 3 is a graph showing the results of a t-test (*<0.05) on the blood D-amino acid ratio (P%D={PD-AA / (PD-AA+L-AA)}×100) of the influenza virus-infected mice group and the non-infected mice (control) group in Example 1. FIG. 3A shows the data for P%D-Ala, FIG. 3B shows the data for P%D-Asn, FIG. 3C shows the data for P%D-Pro, and FIG. 3D shows the data for P%D-Ser. According to these data, the P%D-Ala of the influenza virus-infected mice is significantly lower than that of the control group, and P%D-Asn, P%D-Pro, and P%D-Ser show a tendency to decrease compared to the control group, so it can be seen that viral infection can be determined based on the decrease in the blood D-amino acid ratio. By performing a panel test using multiple P%D-AA, it is possible to make a more accurate determination. By performing a panel test combining the above-mentioned PD-AA and P%D-AA, the accuracy of the determination can be further improved. In addition, these judgment results can be used to verify the validity of test results for other viral infections and / or viral infectious diseases, such as PCR tests.

[0084] 4 is a graph showing the body weight of influenza virus-infected mice in comparison with non-infected mice in Example 1. This data shows that the body weight of the mice, which is one of the symptoms of influenza virus infection, was clearly reduced, indicating that the influenza model mice functioned as severe influenza models.

[0085] [Example 2: Clinical Trials for COVID-19] An observational study was conducted as part of a phase 1 and 2 single-arm, multicenter, open-blind clinical study to evaluate the efficacy and safety of viral adsorption therapy in severe patients with COVID-19 (CA1CH-COVID, Japan Clinical Trial Registration ID: 052200134).

[0086] The inclusion criteria for subjects were 18 years of age or older, positive blood test for SARS-CoV-2 in a pre-registration test, and severe COVID-19 cases requiring either artificial ventilation or ECMO (n=5, including one patient with renal failure). The exclusion criteria were patients with unstable hemodynamics who could not undergo blood purification therapy. The primary outcome was discontinuation of artificial ventilation or ECMO after recovery, and the secondary outcomes were (i) improvement of respiratory function as measured by the ratio of arterial oxygen pressure to inspired oxygen pressure (PaO2 / FiO2 ratio), and (ii) negative blood test for SARS-CoV-2.

[0087] After screening for eligibility, including a blood test for SARS-CoV-2 on day 0, patients underwent blood purification therapy using a SARS-CoV-2 adsorption column for three consecutive days. During the period from days 0 to 15, chiral amino acids (PD-AA, PL-AA) in the plasma of blood collected from the subjects were quantitatively analyzed by HPLC. As a control, a healthy population described in the inventor's previously published publications, E. Clinical. Medicine, 2022, (Non-Patent Document 3) and Sci. Rep., 2019, (Non-Patent Document 8), was used.

[0088] This clinical study was approved by the Osaka University Central Ethics Committee (#CRB5180007), and written informed consent was obtained from all subjects. The study was conducted in accordance with the Declaration of Helsinki and ethical guidelines for medical research involving human subjects.

[0089] FIG. 5 is a graph showing the time course of PD-AA in the treatment of COVID-19 patients in Example 2 (excluding patients with renal failure). FIG. 5A shows data for PD-Ala, FIG. 5B shows data for PD-Asn, FIG. 5C shows data for PD-Pro, and FIG. 5D shows data for PD-Ser. The horizontal line indicates the reference range (95% confidence interval) for healthy controls. As is clear from these data, the PD-AA of all COVID-19 patients is lower than the reference range or shows a tendency to decrease, so that it is possible to determine whether a subject is infected with COVID-19 based on the decrease or tendency to decrease in the amount of D-amino acids in the blood. In addition, the patient group was in a severe state requiring ECMO on day 0, but showed a recovery state when treatment (viral adsorption therapy) was started, and all patients eventually progressed to a state of remission or cure. PD-AA shows a characteristic pattern of decreasing during the acute phase (worsening and severe phase), then increasing during the recovery phase (improvement phase), and finally converging or approaching the reference range, so it is possible to classify the stage of viral infection using the amount of D-amino acids in the blood. It can also be used to evaluate the effectiveness of treatments such as viral adsorption therapy and to assist in the selection of effective treatments.

[0090] FIG. 6 is a graph showing the results of a t-test (*<0.05) on the amount of D-amino acids in the blood of the COVID-19 patient group and the healthy control group in Example 2. FIG. 6A shows data on PD-Ala, FIG. 6B shows data on PD-Asn, FIG. 6C shows data on PD-Pro, and FIG. 6D shows data on PD-Ser. According to these data, PD-Ala, PD-Pro, and PD-Ser in the COVID-19 patient group are significantly lower (decreased) than in the healthy control group, and PD-Asn shows a tendency to decrease compared to the healthy control group, so it can be seen that a viral infection can be determined based on a decrease in the amount of D-amino acids in the blood. By performing a panel test using multiple PD-AAs, it is possible to make a more accurate determination.

[0091] FIG. 7 is a graph showing the results of a t-test (*<0.05) on the blood D-amino acid ratio (%PD={PD-AA / (PD-AA+PL-AA)}×100) of the COVID-19 patient group and the healthy control group in Example 2. FIG. 7A shows the data for P%D-Ala, FIG. 7B shows the data for P%D-Asn, FIG. 7C shows the data for P%D-Pro, and FIG. 7D shows the data for P%D-Ser. As is clear from these data, the P%D-AA in the COVID-19 patient group is significantly lower than that in the healthy control group, so that a viral infection can be determined based on a decrease in the blood D-amino acid ratio. By performing a panel test using multiple P%D-AA and the PD-AA in FIG. 6, a more accurate determination can be made. By performing a panel test combining the above-mentioned PD-AA and P%D-AA, the accuracy of the determination can be further improved. In addition, these determination results can be used to verify the validity of test results for other viral infections and / or viral infections, such as PCR tests.

[0092] FIG. 8 is a graph showing the time course of PD-AA in the treatment course of end-stage renal failure patients who were excluded from the above analysis among the COVID-19 patient group in Example 2. According to this data, the PD-AA of end-stage renal failure patients suffering from COVID-19 has a high baseline as known from Patent Document 5 and Non-Patent Document 9, but shows a characteristic fluctuation pattern similar to that of other COVID-19 patients. Therefore, by correcting with a renal function marker (e.g., PCre, etc.), it is possible to perform evaluation and assessment similar to the examples shown in FIG. 6 and FIG. 7 above.

[0093] FIG. 9 is a receiver operating characteristic (ROC) curve for COVID-19 prediction (diagnosis) using the PD-AA of the subject in Example 2 as a variable. FIG. 9A shows data for PD-Ala, FIG. 9B shows data for PD-Pro, and FIG. 9C shows data for PD-Ser. According to these data, the AUC (area under the curve) when PD-Ala is used as a variable is 0.929, the AUC when PD-Pro is used as a variable is 0.967, and the AUC when PD-Ser is used as a variable is 0.878, showing that all of them have high prediction (diagnosis) ability.

[0094] Figure 10 is an ROC curve for prediction (diagnosis) of COVID-19 using the P%D-AA of the subject in Example 2 as a variable. Figure 10A shows data for P%D-Ala, Figure 10B shows data for P%D-Pro, and Figure 10C shows data for P%D-Ser. These data show that the AUC when P%D-Ala is used as a variable is 1.000, the AUC when P%D-Pro is used as a variable is 1.000, and the AUC when P%D-Ser is used as a variable is 0.996, indicating that all of these have extremely high prediction (diagnosis) capabilities.

[0095] 11 is a table showing the AUC of the ROC curve for prediction (diagnosis) of COVID-19 using multiple PD-AA of a subject as multivariables in Example 2. According to these data, with two variables, the AUC of PD-Ser·Pro is 0.967, the AUC of PD-Ser·Ala is 0.946, and the AUC of PD-Pro·Ala is 0.982, and with three variables, the AUC of PD-Ser·Pro·Ala is 1.000, indicating that all have high prediction (diagnosis) ability. [Industrial Applicability]

[0096] The present invention is extremely useful in the fields of diagnosis and treatment of viral infection and / or viral infectious diseases.

Claims

1. 1. A method for providing information regarding a viral infection and / or viral disease in a subject, comprising: using the subject's D-amino acid indicator to determine whether the subject is infected with a virus and / or ... and providing information about the subject's viral infection and / or viral infectious disease based on the results of the determination. The method includes:

2. The method described in claim 1, wherein the D-amino acid is one or more D-amino acids selected from the group consisting of D-proline, D-serine, D-alanine, and D-asparagine.

3. The method according to claim 1 or 2, wherein the index for D-amino acids is a measured value for D-amino acids in blood, or a corrected value or correction formula thereof.

4. The method according to claim 3, wherein the index for the D-amino acid is a value or formula obtained by correcting the amount of the D-amino acid with an index for a substance in the body of the subject (for example, an L-amino acid).

5. The method according to claim 3, wherein the index related to the D-amino acid is a value or formula obtained by correcting the amount of the D-amino acid by an index related to the renal function of the subject.

6. 3. The method of claim 1 or 2, wherein the virus is a virus belonging to a family selected from orthomyxoviruses, coronaviruses, paramyxoviruses, rhabdoviruses, arenaviruses, bunyaviruses, filoviruses, retroviruses, togaviruses, flaviviruses, picornaviruses, astroviruses, caliciviruses, reoviruses, parvoviruses, adenoviruses, papillomaviruses, polyomaviruses, herpesviruses, hepadnaviruses, and poxviruses.

7. The determination of the viral infection and / or detection of a viral infection is - determining that the subject is infected with a virus and / or suffers from a viral infection when the index related to the D-amino acid of the subject shows a downward trend; 3. The method of claim 1 or 2, comprising:

8. The determination of the viral infection and / or the stage of the viral infection, determining that the subject is in a worsening state of viral infection when the index related to the D-amino acid of the subject is decreased; determining that the subject is in an improved state of viral infection when the index related to the D-amino acid of the subject is increased; and / or - determining that the subject is cured of the viral infection when the index related to the D-amino acid of the subject repeatedly decreases and increases and then converges to a healthy reference range; 3. The method of claim 1 or 2, selected from the group consisting of:

9. The method according to claim 1 or 2, wherein the detection and / or staging of the viral infection and / or viral infectious disease is determined by comparing the subject's indicator for D-amino acids with a determination criterion for viral infection and / or viral infectious disease.

10. The subject's viral infection and / or information regarding the viral infection comprises: - the presence or absence of viral infection and / or viral infection detected in said subject; - staging the subject's viral infection; - verifying the validity of test results and / or diagnostic results for viral infection and / or viral infection in said subject; and -Selection of a treatment method for a viral infection in the subject The method of claim 1 or 2, wherein the information is information about an event selected from the group consisting of:

11. 11. The method of claim 10, wherein the treatment for the viral infection comprises a treatment selected from antiviral drugs, blood purification therapy, a ventilator, and extracorporeal membrane oxygenation (ECMO).

12. A system for carrying out the method according to claim 1 or 2, comprising: The system includes an input unit, an analysis and measurement unit, a memory unit, a data processing unit, and an output unit; The input unit inputs information from a subject, the analyzing and measuring unit analyzes and measures the information from the subject inputted via the input unit to obtain an index related to the D-amino acid of the subject; the storage unit stores a determination criterion for a virus infection and / or a viral infection; the data processing unit processes the indicators of the subject acquired by the analysis and measurement unit based on the judgment criteria stored in the storage unit, thereby determining whether the subject is infected with a virus and / or a viral infectious disease; The output unit outputs the result of the determination by the data processing unit as information regarding the viral infection and / or viral infectious disease of the subject. The system is configured as follows:

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