WO2021107548A1 - Biomarker for diagnosis of infections or infectious diseases caused by nontuberculous mycobacteria - Google Patents

Biomarker for diagnosis of infections or infectious diseases caused by nontuberculous mycobacteria Download PDF

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WO2021107548A1
WO2021107548A1 PCT/KR2020/016658 KR2020016658W WO2021107548A1 WO 2021107548 A1 WO2021107548 A1 WO 2021107548A1 KR 2020016658 W KR2020016658 W KR 2020016658W WO 2021107548 A1 WO2021107548 A1 WO 2021107548A1
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mycobacterium
infection
tuberculous
infectious disease
diagnosing
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French (fr)
Korean (ko)
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신성재
박지해
김크은산
고원중
전병우
김수영
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주식회사 큐라티스
연세대학교 산학협력단
사회복지법인 삼성생명공익재단
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Publication of WO2021107548A1 publication Critical patent/WO2021107548A1/en

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    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/68—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving proteins, peptides or amino acids
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N24/00—Investigating or analyzing materials by the use of nuclear magnetic resonance, electron paramagnetic resonance or other spin effects
    • G01N24/08—Investigating or analyzing materials by the use of nuclear magnetic resonance, electron paramagnetic resonance or other spin effects by using nuclear magnetic resonance
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
    • G01N30/02—Column chromatography
    • G01N30/62—Detectors specially adapted therefor
    • G01N30/72—Mass spectrometers
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
    • G01N30/02—Column chromatography
    • G01N30/88—Integrated analysis systems specially adapted therefor, not covered by a single one of the groups G01N30/04 - G01N30/86
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/68—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving proteins, peptides or amino acids
    • G01N33/6803—General methods of protein analysis not limited to specific proteins or families of proteins
    • G01N33/6848—Methods of protein analysis involving mass spectrometry
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
    • G01N30/02—Column chromatography
    • G01N30/88—Integrated analysis systems specially adapted therefor, not covered by a single one of the groups G01N30/04 - G01N30/86
    • G01N2030/8809—Integrated analysis systems specially adapted therefor, not covered by a single one of the groups G01N30/04 - G01N30/86 analysis specially adapted for the sample
    • G01N2030/8813—Integrated analysis systems specially adapted therefor, not covered by a single one of the groups G01N30/04 - G01N30/86 analysis specially adapted for the sample biological materials
    • G01N2030/8818—Integrated analysis systems specially adapted therefor, not covered by a single one of the groups G01N30/04 - G01N30/86 analysis specially adapted for the sample biological materials involving amino acids
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N2800/00—Detection or diagnosis of diseases
    • G01N2800/26—Infectious diseases, e.g. generalised sepsis

Definitions

  • the present invention relates to a biomarker for diagnosing infection or infectious disease caused by non-tuberculous mycobacteria, and a kit for the diagnosis or a method for providing information for diagnosis.
  • Mycobacterium includes not only species that cause serious diseases in humans and animals, such as tuberculosis, tuberculosis bovine tuberculosis, and leprosy, but also fungal species called opportunistic bacteria, and parasitic objects found in the natural environment. About 72 species such as saprophytic species are known so far, of which 25 are known to be related to human diseases. These Mycobacterium genus are not easily dyed with commonly used dyeing solutions, but once dyed, they are also called acid-fighting bacteria because they are not easily decolorized even when treated with alcohol or hydrochloric acid.
  • Nontuberculous mycobacteria refers to mycobacteria other than Mycobacterium tuberculosis complex and Mycobacterium leprae.
  • MAC Mycobacterium avium complex
  • MAB Mycobacterium abscessus
  • M. abscessus subspecies Absesu. s M. abscessus subspecies abscessus
  • M. abscessus subspecies massiliense M. abscessus subspecies massiliense
  • One object of the present invention is to provide a biomarker composition for diagnosing infection or infectious disease of non-tuberculous mycobacterium.
  • Another object of the present invention is to provide a kit for diagnosing non-tuberculous mycobacterium infection or infectious disease.
  • Another object of the present invention is to provide a method for providing information for diagnosing non-tuberculous mycobacterium infection or infectious disease.
  • biomarker composition for diagnosis of non-tuberculous mycobacterium infection or infectious disease, including metabolites.
  • diagnosis refers to determining the susceptibility of an object, that is, a test subject to a specific disease or disorder, determining whether an object currently has a specific disease or disorder, Determining the prognosis of a subject afflicted with a disease or disorder, or therametrics (eg, monitoring the condition of a subject to provide information about treatment efficacy).
  • the diagnostic biomarker according to the present invention is a material capable of diagnosing a non-tuberculous mycobacterium infection or post-infection disease by distinguishing it from a healthy normal control group, and is a biological biomarker derived from a subject infected with non-tuberculous mycobacteria compared to a normal control group. Metabolites showing an increase or decrease in the sample, preferably blood metabolites.
  • the "metabolite” is also called a metabolite or metabolite, and is an intermediate product or product of metabolism.
  • metabolites are fuel, structure, signaling, catalytic and inhibitory effects on enzymes, their own catalytic activity (usually as cofactors for enzymes), defense, and interactions with other organisms (eg, pigments, aromatic compounds). , pheromones).
  • Primary metabolites are directly involved in normal growth, development and reproduction. Secondary metabolites are not directly involved in these processes, but usually have important ecological functions.
  • the metabolite refers to a sample of biological origin, that is, a metabolite obtained from a biological sample
  • the biological sample refers to a biological body fluid, tissue or cell, for example, whole blood, leukocyte. (leukocytes), peripheral blood mononuclear cells, buffy coat, plasma, serum, sputum, tears, mucus, nasal washes, nasal aspirate, breath, urine, semen, saliva, peritoneal washings, ascites, cystic fluid , meningeal fluid, amniotic fluid, glandular fluid, pancreatic fluid, lymph fluid, pleural fluid, nipple aspirate, bronchial aspirate ( One selected from the group consisting of bronchial aspirate, synovial fluid, joint aspirate, organ secretions, cell, cell extract, and cerebrospinal fluid. It may be more than, but preferably whole blood (whole blood), plasma (plasma) or serum (serum) may be, and more of bronchial
  • whole blood, plasma or serum may be pretreated to detect the metabolite.
  • it may include filtration, distillation, extraction, separation, concentration, inactivation of interfering components, addition of reagents, and the like.
  • the metabolite may include a substance produced by metabolism and metabolic processes or a substance generated by chemical metabolism by biological enzymes and molecules.
  • the metabolite is preferably a metabolite obtained from a liquid sample derived from blood, preferably serum, and specific examples include amino acids, amino acid derivatives, allantoin, N,N- Dimethylglycine (N,N-Dimethylglycine), 2-hydroxyglutaric acid, Choline, Hypoxanthine, Lactate, Malic acid and Glycerol 3- It may include one or more selected from the group consisting of phosphate (Glycerol 3-phosphate).
  • the 2-hydroxyglutaric acid is preferably in the D-form, but is not limited thereto.
  • the lactate is preferably in the S-form, but is not limited thereto.
  • the malic acid is preferably in the L-form, but is not limited thereto.
  • the amino acids and their derivatives are Arginine, Proline, Threonine, Leucine, Lysine, Glutamate, Tryptophan, Asparagine, Methionine (Methionine), serine (Serine), homoserine (Homoserine) and taurine (Taurine) may include one or more selected from the group consisting of.
  • the amino acid may be in L-form, preferably arginine, proline, threonine, leucine, lysine, glutamate.
  • Tryptophan, asparagine, methionine or serine is preferably in the L-form, but is not limited thereto.
  • the non-tuberculous mycobacteria are Mycobacterium avium (M. avium), Mycobacterium abscessus (M. abscessus), Mycobacterium flavesense (M. flavescence), Mycobacterium Rum africanum (M. africanum), Mycobacterium bovis (M. bovis), Mycobacterium cellone (M. chelonae), Mycobacterium cellatum (M. celatum), Mycobacterium portuitum (M. fortuitum), Mycobacterium gordonae (M. gordonae), Mycobacterium gastri (M. gastri), Mycobacterium haemophilum (M.
  • Mycobacterium intracellulare M. intracellulare
  • mycobacterium kansasii M. kansasii
  • mycobacterium malmoens M. malmoense
  • mycobacterium marinum M. marinum
  • mycobacterium sulgai M) szulgai
  • Mycobacterium terrae M. terrae
  • Mycobacterium scrofulaceum M. scrofulaceum
  • Mycobacterium Ulcerans M. ulcerans
  • Mycobacterium simiae M. simiae
  • Mycobacterium xenopi M. xenopi
  • M. xenopi is preferably selected from the group consisting of, but is not limited thereto.
  • the non-tuberculous mycobacterium-infectious disease includes all clinical symptoms caused by the non-tuberculous mycobacterium infection, and the infectious disease is a lung disease, lymphadenitis, skin/soft tissue/bone infection or disseminated disease, etc. may include.
  • the infectious lung disease caused by the non-tuberculous mycobacterium may be a fibrocavitary form, a nodular bronchiectatic form, or a combination thereof.
  • the infectious lung disease may be accompanied by cough, sputum, blood sputum, fever, dyspnea, chest pain, or a combination thereof.
  • the infectious lung disease caused by the non-tuberculous mycobacterium is a microorganism that causes an infectious lung disease, for example, M. avium complex, M. kansasii. ), Mycobacterium abscessus (M. abscessus), Mycobacterium xenopi (M. xenopi), Mycobacterium malmoens (M. malmoense), Mycobacterium fortuitum (M. fortuitum) , Mycobacterium marinum (M. marinum), Mycobacterium cellone (M. chelonae), Mycobacterium gordonae (M. gordonae), Mycobacterium szulgai (M.
  • M. avium complex M. kansasii.
  • Mycobacterium abscessus M. abscessus
  • Mycobacterium xenopi M. xenopi
  • Mycobacterium malmoens M. malmoense
  • Mycobacterium Cobacterium cellatum M. celatum
  • Mycobacterium scrofulaceum M. scrofulaceum
  • Mycobacterium terae M. terrae
  • Mycobacterium simiae M. simiae It may be a lung disease, but is not limited thereto.
  • a kit for diagnosing non-tuberculous mycobacteria infection or infectious disease including a quantitative device for measuring the concentration of a metabolite.
  • the metabolite refers to a sample of biological origin, that is, a metabolite obtained from a biological sample
  • the biological sample refers to a biological body fluid, tissue or cell, for example, whole blood, leukocyte. (leukocytes), peripheral blood mononuclear cells, buffy coat, plasma, serum, sputum, tears, mucus, nasal washes, nasal aspirate, breath, urine, semen, saliva, peritoneal washings, ascites, cystic fluid , meningeal fluid, amniotic fluid, glandular fluid, pancreatic fluid, lymph fluid, pleural fluid, nipple aspirate, bronchial aspirate ( One selected from the group consisting of bronchial aspirate, synovial fluid, joint aspirate, organ secretions, cell, cell extract, and cerebrospinal fluid. It may be more than, but preferably whole blood (whole blood), plasma (plasma) or serum (serum) may be, and more of bronchial
  • whole blood, plasma or serum may be pretreated to detect the metabolite.
  • it may include filtration, distillation, extraction, separation, concentration, inactivation of interfering components, addition of reagents, and the like.
  • the metabolite is preferably a metabolite obtained from a liquid sample derived from blood, preferably serum, and specific examples include amino acids, amino acid derivatives, allantoin, N,N- Dimethylglycine (N,N-Dimethylglycine), 2-hydroxyglutaric acid, Choline, Hypoxanthine, Lactate, Malic acid and Glycerol 3- It may include one or more selected from the group consisting of phosphate (Glycerol 3-phosphate).
  • the 2-hydroxyglutaric acid is preferably in the D-form, but is not limited thereto.
  • the lactate is preferably in the S-form, but is not limited thereto.
  • the malic acid is preferably in the L-form, but is not limited thereto.
  • the amino acids and their derivatives are Arginine, Proline, Threonine, Leucine, Lysine, Glutamate, Tryptophan, Asparagine, Methionine (Methionine), serine (Serine), homoserine (Homoserine) and taurine (Taurine) may include one or more selected from the group consisting of.
  • the amino acid may be in L-form, preferably arginine, proline, threonine, leucine, lysine, glutamate.
  • Tryptophan, asparagine, methionine or serine is preferably in the L-form, but is not limited thereto.
  • the quantitative device may be a nuclear magnetic resonance spectrometer (NMR), chromatography, or mass spectrometer, but is not limited thereto.
  • NMR nuclear magnetic resonance spectrometer
  • chromatography chromatography
  • mass spectrometer but is not limited thereto.
  • Chromatography used in the present invention is high performance liquid chromatography (HPLC), liquid-solid chromatography (Liquid-Solid Chromatography, LSC), paper chromatography (Paper Chromatography, PC), thin layer chromatography (Thin) -Layer Chromatography (TLC), Gas-Solid Chromatography (GSC), Liquid-Liquid Chromatography (LLC), Foam Chromatography (FC), Emulsion Chromatography (Emulsion) Chromatography (EC), Gas-Liquid Chromatography (GLC), Ion Chromatography (IC), Gel Filtration Chromatography (GFC) or Gel Permeation Chromatography (Gel Permeation Chromatography, GPC), but is not limited thereto, and any quantitative chromatography commonly used in the art may be used.
  • HPLC high performance liquid chromatography
  • LSC liquid-solid chromatography
  • PC Paper chromatography
  • TLC thin layer chromatography
  • GSC Gas-Solid Chromatography
  • LLC Liquid
  • the mass spectrometer may use a conventionally known mass spectrometer without any particular limitation, but specifically, for example, a Fourier transform mass spectrometer (FTMS), a Malditope mass spectrometer (MALDI-TOF MS), It may be Q-TOF MS or LTQ-Orbitrap MS, but is not limited thereto.
  • FTMS Fourier transform mass spectrometer
  • MALDI-TOF MS Malditope mass spectrometer
  • Q-TOF MS Q-TOF MS or LTQ-Orbitrap MS, but is not limited thereto.
  • non-tuberculous mycobacterium infectious disease in the kit of the present invention overlaps with that described in the biomarker composition of the present invention, and thus description thereof will be omitted below to avoid excessive congestion of the specification.
  • it relates to a method for providing information for diagnosing infection or infectious disease of non-tuberculous mycobacteria, comprising measuring the expression level of a metabolite in a biological sample isolated from a target subject. .
  • the "target individual” refers to an individual whose infection is uncertain by non-tuberculous mycobacteria, and is highly likely to be infected.
  • the "biological sample” refers to any material, biological fluid, tissue or cell obtained from or derived from an individual, and includes whole blood, leukocytes, and peripheral blood mononuclear cells.
  • a step of pre-treating the biological sample preferably whole blood, plasma or serum may be performed prior to measuring the expression level of the metabolite.
  • the pretreatment may include, for example, filtration, distillation, extraction, separation, concentration, inactivation of interfering components, addition of reagents, and the like, but is not limited thereto.
  • the metabolite is preferably a metabolite obtained from a liquid sample derived from blood, preferably serum, and specific examples include amino acids, amino acid derivatives, allantoin, N,N- Dimethylglycine (N,N-Dimethylglycine), 2-hydroxyglutaric acid, Choline, Hypoxanthine, Lactate, Malic acid and Glycerol 3- It may include one or more selected from the group consisting of phosphate (Glycerol 3-phosphate).
  • the 2-hydroxyglutaric acid is preferably in the D-form, but is not limited thereto.
  • the lactate is preferably in the S-form, but is not limited thereto.
  • the malic acid is preferably in the L-form, but is not limited thereto.
  • the amino acids and their derivatives are Arginine, Proline, Threonine, Leucine, Lysine, Glutamate, Tryptophan, Asparagine, Methionine (Methionine), serine (Serine), homoserine (Homoserine) and taurine (Taurine) may include one or more selected from the group consisting of.
  • the amino acid may be (L)-form ((L)-form), preferably arginine, proline, threonine, leucine, lysine, Glutamate, tryptophan, asparagine, methionine, or serine may be in (L)-form ((L)-form), but is not limited thereto.
  • the expression level of the metabolite may be performed using a quantitative device.
  • the quantitative device may be a nuclear magnetic resonance spectrometer (NMR), chromatography, or mass spectrometer, but is not limited thereto.
  • Chromatography used in the present invention is high performance liquid chromatography (HPLC), liquid-solid chromatography (Liquid-Solid Chromatography, LSC), paper chromatography (Paper Chromatography, PC), thin layer chromatography (Thin) -Layer Chromatography (TLC), Gas-Solid Chromatography (GSC), Liquid-Liquid Chromatography (LLC), Foam Chromatography (FC), Emulsion Chromatography (Emulsion) Chromatography (EC), Gas-Liquid Chromatography (GLC), Ion Chromatography (IC), Gel Filtration Chromatography (GFC) or Gel Permeation Chromatography (Gel Permeation Chromatography, GPC), but is not limited thereto, and any quantitative chromatography commonly used in the art may be used.
  • HPLC high performance liquid chromatography
  • LSC liquid-solid chromatography
  • PC Paper chromatography
  • TLC thin layer chromatography
  • GSC Gas-Solid Chromatography
  • LLC Liquid
  • the mass spectrometer may use a conventionally known mass spectrometer without any particular limitation, but specifically, for example, a Fourier transform mass spectrometer (FTMS), a Malditope mass spectrometer (MALDI-TOF MS), It may be Q-TOF MS or LTQ-Orbitrap MS, but is not limited thereto.
  • FTMS Fourier transform mass spectrometer
  • MALDI-TOF MS Malditope mass spectrometer
  • Q-TOF MS Q-TOF MS or LTQ-Orbitrap MS, but is not limited thereto.
  • the expression level of the metabolite measured with respect to the biological sample of the subject of interest is increased or decreased compared to the control, it is infected by a non-tuberculous mycobacterium or has a high probability of infection, or an infectious disease has occurred, or It may further include the step of predicting that the likelihood of the onset is high.
  • the method may further include predicting that the patient is infected or has a high probability of being infected, or that an infectious disease has occurred or is highly likely to develop.
  • proline, threonine, leucine, lysine, glutamate, tryptophan measured with respect to the biological sample of the subject of the present invention in the present invention asparagine, methionine, serine, homoserine, taurine, N,N-dimethylglycine (N,N-Dimethylglycine), 2-hydroxyglutaric acid (2 -hydroxyglutaric acid), choline (Choline), hypoxanthine (Hypoxanthine), lactate (Lactate), malic acid (Malic acid) and glycerol 3-phosphate (Glycerol 3-phosphate) one or more expression levels selected from the group consisting of a control group If it is reduced compared to the non-tuberculous mycobacteria, it may further include the step of predicting that the infection or high probability of infection, or the onset of an infectious disease or a high probability of onset.
  • control group is a normal control that is not infected with a non-tuberculous mycobacterium, or a non-tuberculous mycobacterium, or the median value of a patient population that has or has a high probability of infection ( or the average value of the patient).
  • the target subject when infected or highly likely to be infected by a non-tuberculous mycobacterium as described above, or an infectious disease has occurred or is predicted to be highly likely to occur, the target subject is treated as a drug for the disease. , for example, may further include the step of performing an appropriate treatment such as administration of antibiotics.
  • the "antibiotic” may be rifampin, isoniazid, ethambutol, pyrazinamide (PZA), quinolone, or aminoglycoside, but is not limited thereto.
  • the quinolone antibiotic is nalidixic acid, marbofloxacin, oxolinic acid, moxifloxacin, trovafloxacin, gatifloxacin, Flumequine, prulifloxacin, gemifloxacin, ciprofloxacin, sitafloxacin, or clinafloxacin, etc., but may not be limited thereto.
  • aminoglycoside antibiotics include streptomycin, neomycin, framycetin, gentamycin, novobiocin, kanamycin, and amica. It may be syn (amikacin), sisomycin (sisomycin) or spectinomycin (spectinomycin), but is not limited thereto.
  • the method of the present invention it is possible to diagnose whether or not infection is caused by non-tuberculous mycobacteria, and the course of the disease, prognosis, or therapeutic effect can also be tracked.
  • non-tuberculous mycobacterium infectious disease overlaps with that described in the biomarker composition of the present invention, and thus description thereof will be omitted below to avoid excessive congestion of the specification.
  • the present invention by measuring the expression level of a blood metabolite with respect to a biological sample of a target individual, the occurrence or possibility of an infectious disease caused by a non-tuberculous mycobacterium can be simply, easily and accurately predicted.
  • L-Arginine L-arginine
  • MAC Mycobacterium avium complex
  • Figure 2 shows a graph comparing the expression level of allantoin (Allantoin) in serum samples of patients with Mycobacterium avium complex (MAC)-infected lung disease and healthy people before antibiotic treatment in an embodiment of the present invention.
  • Allantoin allantoin
  • L-Proline L-proline
  • MAC Mycobacterium avium complex
  • Figure 4 is a graph comparing the expression level of L-threonine (L-Threonine) in serum samples of patients with Mycobacterium avium complex (MAC)-infected lung disease and healthy people before antibiotic treatment in an embodiment of the present invention. it has been shown
  • FIG. 5 is a graph comparing the expression level of L-leucine in serum samples of Mycobacterium avium complex (MAC)-infected lung disease patients and healthy people before antibiotic treatment in one embodiment of the present invention. it has been shown
  • L-lysine L-Lycine
  • MAC Mycobacterium avium complex
  • L-glutamate L-glutamate
  • MAC Mycobacterium avium complex
  • L-tryptophan L-tryptophan
  • MAC Mycobacterium avium complex
  • Figure 9 shows the expression level of N,N-dimethylglycine (N,N-Dimethylglycine) in serum samples from Mycobacterium avium complex (MAC)-infected lung disease patients and healthy people before antibiotic treatment in an embodiment of the present invention; A graph comparing them is shown.
  • N,N-dimethylglycine N,N-Dimethylglycine
  • FIG. 10 shows a graph comparing the expression level of Homoserine in serum samples of Mycobacterium avium complex (MAC)-infected lung disease patients and healthy people before antibiotic treatment in an embodiment of the present invention.
  • MAC Mycobacterium avium complex
  • FIG. 11 shows D-2-hydroxyglutaric acid in serum samples from patients with Mycobacterium avium complex (MAC)-infected lung disease and healthy people before antibiotic treatment in one embodiment of the present invention; ) shows a graph comparing the expression level.
  • MAC Mycobacterium avium complex
  • L-Asparagine L-asparagine
  • MAC Mycobacterium avium complex
  • L-Methionine L-methionine
  • MAC Mycobacterium avium complex
  • FIG. 14 shows a graph comparing the expression level of choline in serum samples of patients with Mycobacterium avium complex (MAC)-infected lung disease and healthy people before antibiotic treatment in an embodiment of the present invention.
  • MAC Mycobacterium avium complex
  • MAC 15 shows a graph comparing the expression level of hypoxanthine in serum samples of Mycobacterium avium complex (MAC)-infected lung disease patients and healthy people before antibiotic treatment in an embodiment of the present invention.
  • MAC Mycobacterium avium complex
  • S-Lactate S-lactate
  • MAC Mycobacterium avium complex
  • 17 is a graph comparing the expression level of L-malic acid in serum samples of patients with Mycobacterium avium complex (MAC)-infected lung disease and healthy people before antibiotic treatment in one embodiment of the present invention.
  • MAC Mycobacterium avium complex
  • FIG. 19 shows a graph comparing the expression level of taurine in serum samples of patients with Mycobacterium avium complex (MAC)-infected lung disease and healthy people before antibiotic treatment in an embodiment of the present invention.
  • MAC Mycobacterium avium complex
  • L-Serine L-serine
  • MAC Mycobacterium avium complex
  • the present invention relates to a method for providing information for diagnosing infection or infectious disease of non-tuberculous mycobacteria, comprising measuring the expression level of a metabolite in a biological sample isolated from a target subject.
  • the metabolite is preferably a metabolite obtained from a liquid sample derived from blood, preferably serum, and specific examples include amino acids, amino acid derivatives, allantoin, N,N- Dimethylglycine (N,N-Dimethylglycine), 2-hydroxyglutaric acid, Choline, Hypoxanthine, Lactate, Malic acid and Glycerol 3- It may include one or more selected from the group consisting of phosphate (Glycerol 3-phosphate).
  • the amino acid and its derivatives are Arginine, Proline, Threonine, Leucine, Lysine, Glutamate, Tryptophan, Asparagine, Methionine ), serine (Serine), homoserine (Homoserine) and taurine (Taurine) may include one or more selected from the group consisting of.
  • the 2-hydroxyglutaric acid is D-form
  • the lactate is S-form
  • the malic acid Malic acid
  • the amino acid may be in the (L)-form ((L)-form).
  • Mycobacterium avium complex ( avium : 46, intracellulare : 50, total 96) infected lungs collected at Samsung Hospital in Seoul for approximately 6 years from January 2012 to August 2016 96 sera samples from diseased patients before antibiotic treatment and 30 sera from healthy people were prepared.
  • 66 patients with the Mycobacterium avium complex-infected lung disease 66 patients had nodular bronchiectatic form and 30 patients had upper lobe cavitary form.
  • sample quality control SQC
  • SQC sample quality control
  • HC represents the expression level of each metabolite in serum samples of 30 healthy people
  • Dx0 Tx0 (Success & Fail)
  • MAC Mycobacterium avium complex
  • the present invention relates to a biomarker for diagnosing infection or infectious disease caused by non-tuberculous mycobacteria, and a kit for the diagnosis or a method for providing information for diagnosis.

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Abstract

The present invention relates to a method for diagnosing infections or infectious diseases caused by nontuberculous mycobacteria by using metabolites.

Description

비결핵 항산균에 의한 감염 또는 감염 질환의 진단용 바이오마커Biomarker for diagnosis of infection or infectious disease caused by non-tuberculous mycobacteria
본 발명은 비결핵 항산균에 의한 감염 또는 감염 질환을 진단하기 위한 바이오마커와, 상기 진단을 위한 키트 또는 진단을 위한 정보 제공 방법에 관한 것이다. The present invention relates to a biomarker for diagnosing infection or infectious disease caused by non-tuberculous mycobacteria, and a kit for the diagnosis or a method for providing information for diagnosis.
마이코박테리움 (Mycobacterium) 속에는 결핵, 우형결핵, 나병과 같이 사람과 동물에 심각한 질병을 일으키는 균 종 (species)뿐 아니라, 기회 감염균으로 일컬어지는 균 종, 그리고 자연환경에서 볼 수 있는 사물 기생의 균 종 (saprophytic species) 등 현재까지 약 72 종(species)이 알려져 있으며, 그 중 인체 질환과 관련된 것이 25종에 이르는 것으로 알려져 있다. 이러한 마이코박테리움 속은 일반적으로 사용되는 염색액으로는 용이하게 염색되지 않지만 일단 염색되면 알코올이나 염산 등으로 처리시에도 용이하게 탈색되지 않기 때문에 항산균이라고도 불린다.Mycobacterium includes not only species that cause serious diseases in humans and animals, such as tuberculosis, tuberculosis bovine tuberculosis, and leprosy, but also fungal species called opportunistic bacteria, and parasitic objects found in the natural environment. About 72 species such as saprophytic species are known so far, of which 25 are known to be related to human diseases. These Mycobacterium genus are not easily dyed with commonly used dyeing solutions, but once dyed, they are also called acid-fighting bacteria because they are not easily decolorized even when treated with alcohol or hydrochloric acid.
비결핵 항산균(Nontuberculous mycobacteria; NTM)은 결핵균(Mycobacterium tuberculosis complex) 및 나균(Mycobacterium leprae)을 제외한 항산균을 의미한다. 한편, 마이코박테리움 아비움 복합체(Mycobacterium avium complex; MAC)에 속하는 비결핵 항산균주 중 흔히 인간에게서 폐 질환을 일으키는 균주로는 공식적으로 대략 180 종 이상이 규명되었다. MAC는 주로 M. 아비움(M. avium)과 M. 인트라셀룰라(M. intracellulare)를 포함하고, 마이코박테리움 압세수스(Mycobacterium abscessus; MAB)는 주로 M. 압세수스 아종인 압세수스(M. abscessus subspecies abscessus)와 M. 압세수스 아종인 마실리엔시스(M. abscessus subspecies massiliense)를 포함한다. 최근 전세계적으로 비결핵 항산균에 기인한 폐 감염 보고가 증가하고 있지만, 건강한 개체군으로부터 비결핵 항산균 폐 감염 질환자를 구별하기 위한 바이오마커나, 질환에 대한 병태 생리의 연구가 부족한 실정이다.Nontuberculous mycobacteria (NTM) refers to mycobacteria other than Mycobacterium tuberculosis complex and Mycobacterium leprae. Meanwhile, among non-tuberculous mycobacteria belonging to the Mycobacterium avium complex (MAC), more than 180 strains have been officially identified as strains that commonly cause lung disease in humans. MAC mainly contains M. avium and M. intracellulare, and Mycobacterium abscessus (MAB) mainly contains M. abscessus subspecies Absesu. s (M. abscessus subspecies abscessus) and M. abscessus subspecies massiliense (M. abscessus subspecies massiliense). Recently, reports of lung infections caused by non-tuberculous mycobacteria have been increasing worldwide, but biomarkers for differentiating non-tuberculous mycobacterial lung infections from healthy populations or pathophysiology studies of diseases are lacking.
본 발명의 일 목적은 비결핵 항산균의 감염 또는 감염 질환을 진단하기 위한 바이오마커 조성물을 제공하고자 한다. One object of the present invention is to provide a biomarker composition for diagnosing infection or infectious disease of non-tuberculous mycobacterium.
본 발명의 다른 목적은 비결핵 항산균의 감염 또는 감염 질환을 진단하기 위한 키트를 제공하고자 한다.Another object of the present invention is to provide a kit for diagnosing non-tuberculous mycobacterium infection or infectious disease.
본 발명의 또 다른 목적은 비결핵 항산균의 감염 또는 감염 질환을 진단하기 위한 정보 제공 방법을 제공하고자 한다. Another object of the present invention is to provide a method for providing information for diagnosing non-tuberculous mycobacterium infection or infectious disease.
그러나 본 발명이 이루고자 하는 기술적 과제는 이상에서 언급한 과제에 제한되지 않으며, 언급되지 않은 또 다른 과제들은 아래의 기재로부터 당 업계에서 통상의 지식을 가진 자에게 명확하게 이해될 수 있을 것이다.However, the technical task to be achieved by the present invention is not limited to the tasks mentioned above, and other tasks not mentioned will be clearly understood by those of ordinary skill in the art from the following description.
본 발명의 일 구현 예에 따르면, 대사체를 포함하는, 비결핵 항산균의 감염 또는 감염 질환의 진단용 바이오마커 조성물에 관한 것이다. According to one embodiment of the present invention, it relates to a biomarker composition for diagnosis of non-tuberculous mycobacterium infection or infectious disease, including metabolites.
본 발명의 명세서에서 용어 "진단"은 특정 질병 또는 질환에 대한 한 객체 즉 검사 대상자의 감수성(susceptibility)을 판정하는 것, 한 객체가 특정 질병 또는 질환을 현재 가지고 있는 지 여부를 판정하는 것, 특정 질병 또는 질환에 걸린 한 객체의 예후(prognosis)를 판정하는 것 또는 테라메트릭스(therametrics)(예컨대, 치료 효능에 대한 정보를 제공하기 위하여 객체의 상태를 모니터링 하는 것)을 포함한다. As used herein, the term “diagnosis” refers to determining the susceptibility of an object, that is, a test subject to a specific disease or disorder, determining whether an object currently has a specific disease or disorder, Determining the prognosis of a subject afflicted with a disease or disorder, or therametrics (eg, monitoring the condition of a subject to provide information about treatment efficacy).
본 발명에 따른 진단용 바이오마커는 비결핵 항산균의 감염자 또는 감염 후 질환이 발병한 자를 건강한 정상 대조군과 구분하여 진단할 수 있는 물질로, 정상 대조군에 비하여 비결핵 항산균에 의해 감염된 대상체 유래의 생물학적 시료에서 증가 또는 감소 양상을 보이는 대사체, 바람직하게는 혈액 대사체를 포함한다.The diagnostic biomarker according to the present invention is a material capable of diagnosing a non-tuberculous mycobacterium infection or post-infection disease by distinguishing it from a healthy normal control group, and is a biological biomarker derived from a subject infected with non-tuberculous mycobacteria compared to a normal control group. Metabolites showing an increase or decrease in the sample, preferably blood metabolites.
본 발명에서 상기 "대사체(metabolite)"는 대사물질 또는 대사산물이라고도 불리우며, 물질 대사의 중간 생성물 또는 생성물이다. 이러한 대사체는 연료, 구조, 신호전달, 효소에 대한 촉진 및 저해 효과, 그 자신의 촉매 활성(일반적으로 효소에 대한 보조 인자로서), 방어, 다른 생물체와의 상호작용(예: 색소, 방향 화합물, 페로몬)을 포함하는 다양한 기능을 가지고 있다. 1차 대사체는 정상적인 생장, 발생 및 생식에 직접적으로 관여한다. 2차 대사체는 이러한 과정들에 직접적으로 관여하지 않지만, 대개 중요한 생태학적 기능을 가지고 있다. In the present invention, the "metabolite" is also called a metabolite or metabolite, and is an intermediate product or product of metabolism. These metabolites are fuel, structure, signaling, catalytic and inhibitory effects on enzymes, their own catalytic activity (usually as cofactors for enzymes), defense, and interactions with other organisms (eg, pigments, aromatic compounds). , pheromones). Primary metabolites are directly involved in normal growth, development and reproduction. Secondary metabolites are not directly involved in these processes, but usually have important ecological functions.
본 발명에서 상기 대사체는 생체 기원의 시료, 즉 생물학적 시료로부터 수득한 대사 물질을 말하는 것으로, 상기 생물학적 시료는 생물학적 체액, 조직 또는 세포를 의미하는 것으로, 예를 들면, 전혈(whole blood), 백혈구(leukocytes), 말초혈액 단핵 세포(peripheral blood mononuclear cells), 백혈구 연층(buffy coat), 혈장(plasma), 혈청(serum), 객담(sputum), 눈물(tears), 점액(mucus), 세비액(nasal washes), 비강 흡인물(nasal aspirate), 호흡(breath), 소변(urine), 정액(semen), 침(saliva), 복강 세척액(peritoneal washings), 복수(ascites), 낭종액(cystic fluid), 뇌척수막 액(meningeal fluid), 양수(amniotic fluid), 선액(glandular fluid), 췌장액(pancreatic fluid), 림프액(lymph fluid), 흉수(pleural fluid), 유두 흡인물(nipple aspirate), 기관지 흡인물(bronchial aspirate), 활액(synovial fluid), 관절 흡인물(joint aspirate), 기관 분비물(organ secretions), 세포(cell), 세포 추출물(cell extract) 및 뇌척수액(cerebrospinal fluid) 등으로 이루어진 군에서 선택된 1종 이상일 수 있으나, 바람직하게는 전혈(whole blood), 혈장(plasma) 또는 혈청(serum)일 수 있고, 보다 바람직하게는 혈청(serum)일 수 있다. In the present invention, the metabolite refers to a sample of biological origin, that is, a metabolite obtained from a biological sample, and the biological sample refers to a biological body fluid, tissue or cell, for example, whole blood, leukocyte. (leukocytes), peripheral blood mononuclear cells, buffy coat, plasma, serum, sputum, tears, mucus, nasal washes, nasal aspirate, breath, urine, semen, saliva, peritoneal washings, ascites, cystic fluid , meningeal fluid, amniotic fluid, glandular fluid, pancreatic fluid, lymph fluid, pleural fluid, nipple aspirate, bronchial aspirate ( One selected from the group consisting of bronchial aspirate, synovial fluid, joint aspirate, organ secretions, cell, cell extract, and cerebrospinal fluid. It may be more than, but preferably whole blood (whole blood), plasma (plasma) or serum (serum) may be, and more preferably may be serum (serum).
본 발명에서는 상기 대사체를 검출하기 위해 전혈, 혈장 또는 혈청을 전처리할 수 있다. 예를 들어, 여과, 증류, 추출, 분리, 농축, 방해 성분의 불활성화, 시약의 첨가 등을 포함할 수 있다. 또한, 상기 대사체는 대사 및 대사 과정에 의해 생산된 물질 또는 생물학적 효소 및 분자에 의한 화학적 대사작용으로 발생한 물질 등을 포함할 수 있다.In the present invention, whole blood, plasma or serum may be pretreated to detect the metabolite. For example, it may include filtration, distillation, extraction, separation, concentration, inactivation of interfering components, addition of reagents, and the like. In addition, the metabolite may include a substance produced by metabolism and metabolic processes or a substance generated by chemical metabolism by biological enzymes and molecules.
본 발명에서 상기 대사체는 혈액, 바람직하게는 혈청 기원의 액상 시료로부터 수득한 대사물질인 것이 바람직하고, 구체적인 예를 들면, 아미노산(amino acid), 아미노산 유도체, 알란토인(Allantoin), N,N-디메틸글라이신(N,N-Dimethylglycine), 2-하이드록시글루타릭산(2-hydroxyglutaric acid), 콜린(Choline), 하이포잔틴(Hypoxanthine), 락테이트(Lactate), 말산(Malic acid) 및 글리세롤 3-포스페이트(Glycerol 3-phosphate)로 이루어진 군에서 선택된 1종 이상을 포함할 수 있다. In the present invention, the metabolite is preferably a metabolite obtained from a liquid sample derived from blood, preferably serum, and specific examples include amino acids, amino acid derivatives, allantoin, N,N- Dimethylglycine (N,N-Dimethylglycine), 2-hydroxyglutaric acid, Choline, Hypoxanthine, Lactate, Malic acid and Glycerol 3- It may include one or more selected from the group consisting of phosphate (Glycerol 3-phosphate).
본 발명에서 상기 2-하이드록시글루타릭산(2-hydroxyglutaric acid)은 D-형태(D-form)인 것이 바람직하나, 이에 제한되는 것은 아니다. In the present invention, the 2-hydroxyglutaric acid is preferably in the D-form, but is not limited thereto.
본 발명에서 상기 락테이트(Lactate)는 S-형태(S-form)인 것이 바람직하나, 이에 제한되는 것은 아니다. In the present invention, the lactate is preferably in the S-form, but is not limited thereto.
본 발명에서 상기 말산(Malic acid)은 L-형태(L-form)인 것이 바람직하나, 이에 제한되는 것은 아니다. In the present invention, the malic acid is preferably in the L-form, but is not limited thereto.
본 발명에서 상기 아미노산 및 그 유도체는 알기닌(Arginine), 프롤린(Proline), 트레오닌(Threonine), 류신(Leucine), 라이신(Lycine), 글루타메이트(Glutamate), 트립토판(Tryptophan), 아스파라긴(Asparagine), 메티오닌(Methionine), 세린(Serine), 호모세린(Homoserine) 및 타우린(Taurine)으로 이루어진 군에서 선택된 1종 이상을 포함할 수 있다. In the present invention, the amino acids and their derivatives are Arginine, Proline, Threonine, Leucine, Lysine, Glutamate, Tryptophan, Asparagine, Methionine (Methionine), serine (Serine), homoserine (Homoserine) and taurine (Taurine) may include one or more selected from the group consisting of.
본 발명에서 상기 아미노산은 L-형태(L-form)일 수 있고, 바람직하게는 알기닌(Arginine), 프롤린(Proline), 트레오닌(Threonine), 류신(Leucine), 라이신(Lycine), 글루타메이트(Glutamate), 트립토판(Tryptophan), 아스파라긴(Asparagine), 메티오닌(Methionine) 또는 세린(Serine)은 L-형태(L-form)인 것이 바람직하나, 이에 제한되는 것은 아니다. In the present invention, the amino acid may be in L-form, preferably arginine, proline, threonine, leucine, lysine, glutamate. , Tryptophan, asparagine, methionine or serine is preferably in the L-form, but is not limited thereto.
본 발명에서 상기 비결핵 항산균은 마이코박테리움 아비움(M. avium), 마이코박테리움 압세수스(M. abscessus), 마이코박테리움 플라베센스(M. flavescence), 마이코박테리움 아프리카눔(M. africanum), 마이코박테리움 보비스(M. bovis), 마이코박테리움 첼로네(M. chelonae), 마이코박테리움 셀라툼(M. celatum), 마이코박테리움 포르투이툼(M. fortuitum), 마이코박테리움 고르도네(M. gordonae), 마이코박테리움 가스트리(M. gastri), 마이코박테리움 헤모필룸(M. haemophilum), 마이코박테리움 인트라셀루라레(M. intracellulare), 마이코박테리움 칸사시이(M. kansasii), 마이코박테리움 말모엔스(M. malmoense), 마이코박테리움 마리눔(M. marinum), 마이코박테리움 스줄가이(M. szulgai), 마이코박테리움 테레(M. terrae), 마이코박테리움 스크로풀라세움(M. scrofulaceum), 마이코박테리움 울서란스(M. ulcerans), 마이코박테리움 시미애(M. simiae) 및 마이코박테리움 제노피(M. xenopi)로 구성된 군으로부터 선택된 것이 바람직하나, 이에 제한되는 것은 아니다. In the present invention, the non-tuberculous mycobacteria are Mycobacterium avium (M. avium), Mycobacterium abscessus (M. abscessus), Mycobacterium flavesense (M. flavescence), Mycobacterium Rum africanum (M. africanum), Mycobacterium bovis (M. bovis), Mycobacterium cellone (M. chelonae), Mycobacterium cellatum (M. celatum), Mycobacterium portuitum (M. fortuitum), Mycobacterium gordonae (M. gordonae), Mycobacterium gastri (M. gastri), Mycobacterium haemophilum (M. haemophilum), Mycobacterium intracellulare (M. intracellulare), mycobacterium kansasii (M. kansasii), mycobacterium malmoens (M. malmoense), mycobacterium marinum (M. marinum), mycobacterium sulgai (M) szulgai), Mycobacterium terrae (M. terrae), Mycobacterium scrofulaceum (M. scrofulaceum), Mycobacterium Ulcerans (M. ulcerans), Mycobacterium simiae (M. simiae) and Mycobacterium xenopi (M. xenopi) is preferably selected from the group consisting of, but is not limited thereto.
본 발명에서 상기 비결핵 항산균 감염 질환은 상기 비결핵 항산균의 감염에 의해 나타나는 모든 임상적 증상을 포함하는 것으로, 상기 감염 질환은 폐 질환, 림프절염, 피부·연조직·골감염증 또는 파종성 질환 등을 포함할 수 있다.In the present invention, the non-tuberculous mycobacterium-infectious disease includes all clinical symptoms caused by the non-tuberculous mycobacterium infection, and the infectious disease is a lung disease, lymphadenitis, skin/soft tissue/bone infection or disseminated disease, etc. may include.
본 발명에서 상기 비결핵 항산균에 의한 감염성 폐 질환은 섬유공동형 (fibrocavitary form), 기관지 확장증형 (nodular bronchiectatic form), 또는 이들의 조합인 것일 수 있다. 또한, 상기 감염성 폐 질환은 기침, 객담, 혈담, 발열, 호흡곤란, 흉통 또는 이들의 조합을 동반하는 것일 수 있다. In the present invention, the infectious lung disease caused by the non-tuberculous mycobacterium may be a fibrocavitary form, a nodular bronchiectatic form, or a combination thereof. In addition, the infectious lung disease may be accompanied by cough, sputum, blood sputum, fever, dyspnea, chest pain, or a combination thereof.
본 발명에서 상기 비결핵 항산균에 의한 감염성 폐 질환은 감염성 폐 질환을 일으키는 균으로, 예를 들면, 마이코박테리움 아비움 복합체(M. avium complex), 마이코박테리움 칸사시이(M. kansasii), 마이코박테리움 압세수스(M. abscessus), 마이코박테리움 제노피(M. xenopi), 마이코박테리움 말모엔스(M. malmoense), 마이코박테리움 포르투이툼(M. fortuitum), 마이코박테리움 마리눔(M. marinum), 마이코박테리움 첼로네(M. chelonae), 마이코박테리움 고르도네(M. gordonae), 마이코박테리움 스줄가이(M. szulgai), 마이코박테리움 셀라툼(M. celatum), 마이코박테리움 스크로풀라세움(M. scrofulaceum), 마이코박테리움 테레(M. terrae) 또는 마이코박테리움 시미애(M. simiae)에 의한 감염성 폐 질환인 것일 수 있으나, 이에 제한되는 것은 아니다. In the present invention, the infectious lung disease caused by the non-tuberculous mycobacterium is a microorganism that causes an infectious lung disease, for example, M. avium complex, M. kansasii. ), Mycobacterium abscessus (M. abscessus), Mycobacterium xenopi (M. xenopi), Mycobacterium malmoens (M. malmoense), Mycobacterium fortuitum (M. fortuitum) , Mycobacterium marinum (M. marinum), Mycobacterium cellone (M. chelonae), Mycobacterium gordonae (M. gordonae), Mycobacterium szulgai (M. szulgai), Mycobacterium Cobacterium cellatum (M. celatum), Mycobacterium scrofulaceum (M. scrofulaceum), Mycobacterium terae (M. terrae) or Mycobacterium simiae (M. simiae) It may be a lung disease, but is not limited thereto.
본 발명의 다른 구현 예에 따르면, 대사체의 농도를 측정하는 정량 장치를 포함하는, 비결핵 항산균의 감염 또는 감염 질환의 진단용 키트에 관한 것이다. According to another embodiment of the present invention, it relates to a kit for diagnosing non-tuberculous mycobacteria infection or infectious disease, including a quantitative device for measuring the concentration of a metabolite.
본 발명에서 상기 대사체는 생체 기원의 시료, 즉 생물학적 시료로부터 수득한 대사 물질을 말하는 것으로, 상기 생물학적 시료는 생물학적 체액, 조직 또는 세포를 의미하는 것으로, 예를 들면, 전혈(whole blood), 백혈구(leukocytes), 말초혈액 단핵 세포(peripheral blood mononuclear cells), 백혈구 연층(buffy coat), 혈장(plasma), 혈청(serum), 객담(sputum), 눈물(tears), 점액(mucus), 세비액(nasal washes), 비강 흡인물(nasal aspirate), 호흡(breath), 소변(urine), 정액(semen), 침(saliva), 복강 세척액(peritoneal washings), 복수(ascites), 낭종액(cystic fluid), 뇌척수막 액(meningeal fluid), 양수(amniotic fluid), 선액(glandular fluid), 췌장액(pancreatic fluid), 림프액(lymph fluid), 흉수(pleural fluid), 유두 흡인물(nipple aspirate), 기관지 흡인물(bronchial aspirate), 활액(synovial fluid), 관절 흡인물(joint aspirate), 기관 분비물(organ secretions), 세포(cell), 세포 추출물(cell extract) 및 뇌척수액(cerebrospinal fluid) 등으로 이루어진 군에서 선택된 1종 이상일 수 있으나, 바람직하게는 전혈(whole blood), 혈장(plasma) 또는 혈청(serum)일 수 있고, 보다 바람직하게는 혈청(serum)일 수 있다. In the present invention, the metabolite refers to a sample of biological origin, that is, a metabolite obtained from a biological sample, and the biological sample refers to a biological body fluid, tissue or cell, for example, whole blood, leukocyte. (leukocytes), peripheral blood mononuclear cells, buffy coat, plasma, serum, sputum, tears, mucus, nasal washes, nasal aspirate, breath, urine, semen, saliva, peritoneal washings, ascites, cystic fluid , meningeal fluid, amniotic fluid, glandular fluid, pancreatic fluid, lymph fluid, pleural fluid, nipple aspirate, bronchial aspirate ( One selected from the group consisting of bronchial aspirate, synovial fluid, joint aspirate, organ secretions, cell, cell extract, and cerebrospinal fluid. It may be more than, but preferably whole blood (whole blood), plasma (plasma) or serum (serum) may be, and more preferably may be serum (serum).
본 발명에서는 상기 대사체를 검출하기 위해 전혈, 혈장 또는 혈청을 전처리할 수 있다. 예를 들어, 여과, 증류, 추출, 분리, 농축, 방해 성분의 불활성화, 시약의 첨가 등을 포함할 수 있다. In the present invention, whole blood, plasma or serum may be pretreated to detect the metabolite. For example, it may include filtration, distillation, extraction, separation, concentration, inactivation of interfering components, addition of reagents, and the like.
본 발명에서 상기 대사체는 혈액, 바람직하게는 혈청 기원의 액상 시료로부터 수득한 대사물질인 것이 바람직하고, 구체적인 예를 들면, 아미노산(amino acid), 아미노산 유도체, 알란토인(Allantoin), N,N-디메틸글라이신(N,N-Dimethylglycine), 2-하이드록시글루타릭산(2-hydroxyglutaric acid), 콜린(Choline), 하이포잔틴(Hypoxanthine), 락테이트(Lactate), 말산(Malic acid) 및 글리세롤 3-포스페이트(Glycerol 3-phosphate)로 이루어진 군에서 선택된 1종 이상을 포함할 수 있다. In the present invention, the metabolite is preferably a metabolite obtained from a liquid sample derived from blood, preferably serum, and specific examples include amino acids, amino acid derivatives, allantoin, N,N- Dimethylglycine (N,N-Dimethylglycine), 2-hydroxyglutaric acid, Choline, Hypoxanthine, Lactate, Malic acid and Glycerol 3- It may include one or more selected from the group consisting of phosphate (Glycerol 3-phosphate).
본 발명에서 상기 2-하이드록시글루타릭산(2-hydroxyglutaric acid)은 D-형태(D-form)인 것이 바람직하나, 이에 제한되는 것은 아니다. In the present invention, the 2-hydroxyglutaric acid is preferably in the D-form, but is not limited thereto.
본 발명에서 상기 락테이트(Lactate)는 S-형태(S-form)인 것이 바람직하나, 이에 제한되는 것은 아니다. In the present invention, the lactate is preferably in the S-form, but is not limited thereto.
본 발명에서 상기 말산(Malic acid)은 L-형태(L-form)인 것이 바람직하나, 이에 제한되는 것은 아니다. In the present invention, the malic acid is preferably in the L-form, but is not limited thereto.
본 발명에서 상기 아미노산 및 그 유도체는 알기닌(Arginine), 프롤린(Proline), 트레오닌(Threonine), 류신(Leucine), 라이신(Lycine), 글루타메이트(Glutamate), 트립토판(Tryptophan), 아스파라긴(Asparagine), 메티오닌(Methionine), 세린(Serine), 호모세린(Homoserine) 및 타우린(Taurine)으로 이루어진 군에서 선택된 1종 이상을 포함할 수 있다. In the present invention, the amino acids and their derivatives are Arginine, Proline, Threonine, Leucine, Lysine, Glutamate, Tryptophan, Asparagine, Methionine (Methionine), serine (Serine), homoserine (Homoserine) and taurine (Taurine) may include one or more selected from the group consisting of.
본 발명에서 상기 아미노산은 L-형태(L-form)일 수 있고, 바람직하게는 알기닌(Arginine), 프롤린(Proline), 트레오닌(Threonine), 류신(Leucine), 라이신(Lycine), 글루타메이트(Glutamate), 트립토판(Tryptophan), 아스파라긴(Asparagine), 메티오닌(Methionine) 또는 세린(Serine)은 L-형태(L-form)인 것이 바람직하나, 이에 제한되는 것은 아니다. In the present invention, the amino acid may be in L-form, preferably arginine, proline, threonine, leucine, lysine, glutamate. , Tryptophan, asparagine, methionine or serine is preferably in the L-form, but is not limited thereto.
본 발명에서 상기 정량 장치는 핵자기 공명 분광 분석기 (NMR), 크로마토그래피 또는 질량분석기일 수 있으나, 이에 제한되지 않는다.In the present invention, the quantitative device may be a nuclear magnetic resonance spectrometer (NMR), chromatography, or mass spectrometer, but is not limited thereto.
본 발명에서 이용되는 크로마토그래피는 고성능 액체 크로마토그래피(High Performance Liquid Chromatography, HPLC), 액체-고체 크로마토그래피(Liquid-Solid Chromatography, LSC), 종이크로마토그래피(Paper Chromatography, PC), 박층 크로마토그래피(Thin-Layer Chromatography, TLC), 기체-고체 크로마토그래피(Gas-Solid Chromatography, GSC), 액체-액체 크로마토그래피(Liquid-Liquid Chromatography, LLC), 포말 크로마토그래피(Foam Chromatography, FC), 유화 크로마토그래피(Emulsion Chromatography, EC), 기체-액체 크로마토그래피(Gas-Liquid Chromatography, GLC), 이온 크로마토그래피(Ion Chromatography, IC), 겔 여과 크로마토그래피(Gel Filtration Chromatograhy, GFC) 또는 겔 투과 크로마토그래피(Gel Permeation Chromatography, GPC)를 포함하나, 이에 제한되지 않고 당업계에서 통상적으로 사용되는 모든 정량용 크로마토그래피를 사용할 수 있다. Chromatography used in the present invention is high performance liquid chromatography (HPLC), liquid-solid chromatography (Liquid-Solid Chromatography, LSC), paper chromatography (Paper Chromatography, PC), thin layer chromatography (Thin) -Layer Chromatography (TLC), Gas-Solid Chromatography (GSC), Liquid-Liquid Chromatography (LLC), Foam Chromatography (FC), Emulsion Chromatography (Emulsion) Chromatography (EC), Gas-Liquid Chromatography (GLC), Ion Chromatography (IC), Gel Filtration Chromatography (GFC) or Gel Permeation Chromatography (Gel Permeation Chromatography, GPC), but is not limited thereto, and any quantitative chromatography commonly used in the art may be used.
본 발명에서 상기 질량분석기는 특별한 제한없이 종래 공지된 질량 분석기를 이용할 수 있지만, 구체적으로 예를 들면, 푸리에 변환 질량분석기(FTMS, Fourier transform mass spectrometer), 말디토프 질량분석기(MALDI-TOF MS), Q-TOF MS 또는 LTQ-Orbitrap MS일 수 있으나, 이에 제한되는 것은 아니다.In the present invention, the mass spectrometer may use a conventionally known mass spectrometer without any particular limitation, but specifically, for example, a Fourier transform mass spectrometer (FTMS), a Malditope mass spectrometer (MALDI-TOF MS), It may be Q-TOF MS or LTQ-Orbitrap MS, but is not limited thereto.
본 발명의 키트에서 비결핵 항산균의 감염 질환에 관한 정의는 상기 본 발명의 바이오마커 조성물에 기재된 바와 중복되어 명세서의 과도한 혼잡을 피하기 위해 이하 그 기재를 생략한다. The definition of the non-tuberculous mycobacterium infectious disease in the kit of the present invention overlaps with that described in the biomarker composition of the present invention, and thus description thereof will be omitted below to avoid excessive congestion of the specification.
본 발명의 또 다른 구현 예에 따르면, 목적하는 개체로부터 분리된 생물학적 시료에서 대사체의 발현 수준을 측정하는 단계를 포함하는 비결핵 항산균의 감염 또는 감염 질환을 진단하기 위한 정보 제공 방법에 관한 것이다. According to another embodiment of the present invention, it relates to a method for providing information for diagnosing infection or infectious disease of non-tuberculous mycobacteria, comprising measuring the expression level of a metabolite in a biological sample isolated from a target subject. .
본 발명에서 상기 "목적하는 개체"란 비결핵 항산균에 의한 감염 여부가 불확실한 개체로, 감염 가능성이 높은 개체를 의미한다.In the present invention, the "target individual" refers to an individual whose infection is uncertain by non-tuberculous mycobacteria, and is highly likely to be infected.
본 발명에서 상기 "생물학적 시료"는 개체로부터 얻어지거나 개체로부터 유래된 임의의 물질, 생물학적 체액, 조직 또는 세포를 의미하는 것으로, 전혈(whole blood), 백혈구(leukocytes), 말초혈액 단핵 세포(peripheral blood mononuclear cells), 백혈구 연층(buffy coat), 혈장(plasma), 혈청(serum), 객담(sputum), 눈물(tears), 점액(mucus), 세비액(nasal washes), 비강 흡인물(nasal aspirate), 호흡(breath), 소변(urine), 정액(semen), 침(saliva), 복강 세척액(peritoneal washings), 복수(ascites), 낭종액(cystic fluid), 뇌척수막 액(meningeal fluid), 양수(amniotic fluid), 선액(glandular fluid), 췌장액(pancreatic fluid), 림프액(lymph fluid), 흉수(pleural fluid), 유두 흡인물(nipple aspirate), 기관지 흡인물(bronchial aspirate), 활액(synovial fluid), 관절 흡인물(joint aspirate), 기관 분비물(organ secretions), 세포(cell), 세포 추출물(cell extract) 및 뇌척수액(cerebrospinal fluid) 등으로 이루어진 군에서 선택된 1종 이상일 수 있으며, 바람직하게는 전혈(whole blood), 혈장(plasma) 또는 혈청(serum)일 수 있고, 보다 바람직하게는 혈청(serum)일 수 있다. In the present invention, the "biological sample" refers to any material, biological fluid, tissue or cell obtained from or derived from an individual, and includes whole blood, leukocytes, and peripheral blood mononuclear cells. mononuclear cells, buffy coat, plasma, serum, sputum, tears, mucus, nasal washes, nasal aspirate , breath, urine, semen, saliva, peritoneal washings, ascites, cystic fluid, meningeal fluid, amniotic fluid fluid, glandular fluid, pancreatic fluid, lymph fluid, pleural fluid, nipple aspirate, bronchial aspirate, synovial fluid, joint It may be one or more selected from the group consisting of joint aspirate, organ secretions, cells, cell extract, and cerebrospinal fluid, and preferably whole blood (whole blood). ), plasma or serum, and more preferably, serum.
본 발명에서는 상기 대사체의 발현 수준을 측정하기에 앞서, 상기 생물학적 시료, 바람직하게는 전혈, 혈장 또는 혈청을 전처리하는 단계를 수행할 수 있다. 본 발명에서 상기 전처리로는, 예를 들어, 여과, 증류, 추출, 분리, 농축, 방해 성분의 불활성화, 시약의 첨가 등을 포함할 수 있으나, 이에 제한되는 것은 아니다. In the present invention, prior to measuring the expression level of the metabolite, a step of pre-treating the biological sample, preferably whole blood, plasma or serum may be performed. In the present invention, the pretreatment may include, for example, filtration, distillation, extraction, separation, concentration, inactivation of interfering components, addition of reagents, and the like, but is not limited thereto.
본 발명에서 상기 대사체는 혈액, 바람직하게는 혈청 기원의 액상 시료로부터 수득한 대사물질인 것이 바람직하고, 구체적인 예를 들면, 아미노산(amino acid), 아미노산 유도체, 알란토인(Allantoin), N,N-디메틸글라이신(N,N-Dimethylglycine), 2-하이드록시글루타릭산(2-hydroxyglutaric acid), 콜린(Choline), 하이포잔틴(Hypoxanthine), 락테이트(Lactate), 말산(Malic acid) 및 글리세롤 3-포스페이트(Glycerol 3-phosphate)로 이루어진 군에서 선택된 1종 이상을 포함할 수 있다. In the present invention, the metabolite is preferably a metabolite obtained from a liquid sample derived from blood, preferably serum, and specific examples include amino acids, amino acid derivatives, allantoin, N,N- Dimethylglycine (N,N-Dimethylglycine), 2-hydroxyglutaric acid, Choline, Hypoxanthine, Lactate, Malic acid and Glycerol 3- It may include one or more selected from the group consisting of phosphate (Glycerol 3-phosphate).
본 발명에서 상기 2-하이드록시글루타릭산(2-hydroxyglutaric acid)은 D-형태(D-form)인 것이 바람직하나, 이에 제한되는 것은 아니다. In the present invention, the 2-hydroxyglutaric acid is preferably in the D-form, but is not limited thereto.
본 발명에서 상기 락테이트(Lactate)는 S-형태(S-form)인 것이 바람직하나, 이에 제한되는 것은 아니다. In the present invention, the lactate is preferably in the S-form, but is not limited thereto.
본 발명에서 상기 말산(Malic acid)은 L-형태(L-form)인 것이 바람직하나, 이에 제한되는 것은 아니다. In the present invention, the malic acid is preferably in the L-form, but is not limited thereto.
본 발명에서 상기 아미노산 및 그 유도체는 알기닌(Arginine), 프롤린(Proline), 트레오닌(Threonine), 류신(Leucine), 라이신(Lycine), 글루타메이트(Glutamate), 트립토판(Tryptophan), 아스파라긴(Asparagine), 메티오닌(Methionine), 세린(Serine), 호모세린(Homoserine) 및 타우린(Taurine)으로 이루어진 군에서 선택된 1종 이상을 포함할 수 있다. In the present invention, the amino acids and their derivatives are Arginine, Proline, Threonine, Leucine, Lysine, Glutamate, Tryptophan, Asparagine, Methionine (Methionine), serine (Serine), homoserine (Homoserine) and taurine (Taurine) may include one or more selected from the group consisting of.
본 발명에서 상기 아미노산은 (L)-형태((L)-form)일 수 있고, 바람직하게는 알기닌(Arginine), 프롤린(Proline), 트레오닌(Threonine), 류신(Leucine), 라이신(Lycine), 글루타메이트(Glutamate), 트립토판(Tryptophan), 아스파라긴(Asparagine), 메티오닌(Methionine) 또는 세린(Serine)은 (L)-형태((L)-form)일 수 있으나, 이에 제한되는 것은 아니다. In the present invention, the amino acid may be (L)-form ((L)-form), preferably arginine, proline, threonine, leucine, lysine, Glutamate, tryptophan, asparagine, methionine, or serine may be in (L)-form ((L)-form), but is not limited thereto.
본 발명에서 상기 대사체의 발현 수준은 정량 장치를 이용하여 수행될 수 있다. 본 발명에서 상기 정량 장치는 핵자기 공명 분광 분석기 (NMR), 크로마토그래피 또는 질량분석기일 수 있으나, 이에 제한되지 않는다.In the present invention, the expression level of the metabolite may be performed using a quantitative device. In the present invention, the quantitative device may be a nuclear magnetic resonance spectrometer (NMR), chromatography, or mass spectrometer, but is not limited thereto.
본 발명에서 이용되는 크로마토그래피는 고성능 액체 크로마토그래피(High Performance Liquid Chromatography, HPLC), 액체-고체 크로마토그래피(Liquid-Solid Chromatography, LSC), 종이크로마토그래피(Paper Chromatography, PC), 박층 크로마토그래피(Thin-Layer Chromatography, TLC), 기체-고체 크로마토그래피(Gas-Solid Chromatography, GSC), 액체-액체 크로마토그래피(Liquid-Liquid Chromatography, LLC), 포말 크로마토그래피(Foam Chromatography, FC), 유화 크로마토그래피(Emulsion Chromatography, EC), 기체-액체 크로마토그래피(Gas-Liquid Chromatography, GLC), 이온 크로마토그래피(Ion Chromatography, IC), 겔 여과 크로마토그래피(Gel Filtration Chromatograhy, GFC) 또는 겔 투과 크로마토그래피(Gel Permeation Chromatography, GPC)를 포함하나, 이에 제한되지 않고 당업계에서 통상적으로 사용되는 모든 정량용 크로마토그래피를 사용할 수 있다. Chromatography used in the present invention is high performance liquid chromatography (HPLC), liquid-solid chromatography (Liquid-Solid Chromatography, LSC), paper chromatography (Paper Chromatography, PC), thin layer chromatography (Thin) -Layer Chromatography (TLC), Gas-Solid Chromatography (GSC), Liquid-Liquid Chromatography (LLC), Foam Chromatography (FC), Emulsion Chromatography (Emulsion) Chromatography (EC), Gas-Liquid Chromatography (GLC), Ion Chromatography (IC), Gel Filtration Chromatography (GFC) or Gel Permeation Chromatography (Gel Permeation Chromatography, GPC), but is not limited thereto, and any quantitative chromatography commonly used in the art may be used.
본 발명에서 상기 질량분석기는 특별한 제한없이 종래 공지된 질량 분석기를 이용할 수 있지만, 구체적으로 예를 들면, 푸리에 변환 질량분석기(FTMS, Fourier transform mass spectrometer), 말디토프 질량분석기(MALDI-TOF MS), Q-TOF MS 또는 LTQ-Orbitrap MS일 수 있으나, 이에 제한되는 것은 아니다.In the present invention, the mass spectrometer may use a conventionally known mass spectrometer without any particular limitation, but specifically, for example, a Fourier transform mass spectrometer (FTMS), a Malditope mass spectrometer (MALDI-TOF MS), It may be Q-TOF MS or LTQ-Orbitrap MS, but is not limited thereto.
본 발명에서 상기 목적하는 개체의 생물학적 시료에 대하여 측정된 상기 대사체의 발현 수준이 대조군에 비하여 증가 또는 감소된 경우, 비결핵 항산균에 의해 감염되었거나 감염 가능성이 높거나, 또는 감염 질환이 발병하였거나 발병 가능성이 높은 것으로 예측하는 단계를 더 포함할 수 있다.In the present invention, when the expression level of the metabolite measured with respect to the biological sample of the subject of interest is increased or decreased compared to the control, it is infected by a non-tuberculous mycobacterium or has a high probability of infection, or an infectious disease has occurred, or It may further include the step of predicting that the likelihood of the onset is high.
본 발명의 일 예시로, 본 발명에서 상기 목적하는 개체의 생물학적 시료에 대하여 측정된 알기닌(Arginine) 및 알란토인(Allantoin) 중 적어도 하나의 발현 수준이 대조군에 비하여 증가된 경우, 비결핵 항산균에 의해 감염되었거나 감염 가능성이 높거나, 또는 감염 질환이 발병하였거나 발병 가능성이 높은 것으로 예측하는 단계를 더 포함할 수 있다.As an example of the present invention, when the expression level of at least one of arginine and allantoin measured with respect to the biological sample of the subject of the present invention is increased compared to the control, by non-tuberculous mycobacteria The method may further include predicting that the patient is infected or has a high probability of being infected, or that an infectious disease has occurred or is highly likely to develop.
본 발명의 다른 예시로, 본 발명에서 상기 목적하는 개체의 생물학적 시료에 대하여 측정된 프롤린(Proline), 트레오닌(Threonine), 류신(Leucine), 라이신(Lycine), 글루타메이트(Glutamate), 트립토판(Tryptophan), 아스파라긴(Asparagine), 메티오닌(Methionine), 세린(Serine), 호모세린(Homoserine), 타우린(Taurine), N,N-디메틸글라이신(N,N-Dimethylglycine), 2-하이드록시글루타릭산(2-hydroxyglutaric acid), 콜린(Choline), 하이포잔틴(Hypoxanthine), 락테이트(Lactate), 말산(Malic acid) 및 글리세롤 3-포스페이트(Glycerol 3-phosphate)으로 이루어진 군에서 선택된 1종 이상의 발현 수준이 대조군에 비하여 감소된 경우, 비결핵 항산균에 의해 감염되었거나 감염 가능성이 높거나, 또는 감염 질환이 발병하였거나 발병 가능성이 높은 것으로 예측하는 단계를 더 포함할 수 있다.As another example of the present invention, proline, threonine, leucine, lysine, glutamate, tryptophan measured with respect to the biological sample of the subject of the present invention in the present invention , asparagine, methionine, serine, homoserine, taurine, N,N-dimethylglycine (N,N-Dimethylglycine), 2-hydroxyglutaric acid (2 -hydroxyglutaric acid), choline (Choline), hypoxanthine (Hypoxanthine), lactate (Lactate), malic acid (Malic acid) and glycerol 3-phosphate (Glycerol 3-phosphate) one or more expression levels selected from the group consisting of a control group If it is reduced compared to the non-tuberculous mycobacteria, it may further include the step of predicting that the infection or high probability of infection, or the onset of an infectious disease or a high probability of onset.
본 발명에서 상기 "대조군"이란 비결핵 항산균에 의해 감염되지 않은 정상 대조군이거나, 비결핵 항산균에 의해 감염되었거나 감염 가능성이 높거나, 또는 감염 질환이 발병하였거나 발병 가능성이 높은 환자 모집단의 중앙값(또는 해당 환자의 평균값)일 수 있다.In the present invention, the "control group" is a normal control that is not infected with a non-tuberculous mycobacterium, or a non-tuberculous mycobacterium, or the median value of a patient population that has or has a high probability of infection ( or the average value of the patient).
더 나아가, 본 발명에서 상기와 같이 비결핵 항산균에 의해 감염되었거나 감염 가능성이 높거나, 또는 감염 질환이 발병하였거나 발병 가능성이 높은 것으로 예측되는 경우, 상기 목적하는 개체에 대하여 그 질환에 대한 약제로, 예를 들면 항생제 투여 등 적절한 치료를 수행하는 단계를 추가로 포함할 수 있다. Furthermore, in the present invention, when infected or highly likely to be infected by a non-tuberculous mycobacterium as described above, or an infectious disease has occurred or is predicted to be highly likely to occur, the target subject is treated as a drug for the disease. , for example, may further include the step of performing an appropriate treatment such as administration of antibiotics.
본 발명에서 상기 "항생제"는 리팜핀, 아이소니아지드, 에탐부톨, 피라지나마이드(pyrazinamide, PZA), 퀴놀론계, 또는 아미노글라이코사이드계일 수 있으나, 이에 제한되는 것은 아니다. 여기서 상기 퀴놀론계 항생제는 날리딕스산(nalidixic acid), 마보플록사신(marbofloxacin), 옥솔린산(oxolinic acid), 목시플록사신(moxifloxacin), 트로바플록사신(trovafloxacin), 가티플록사신(gatifloxacin), 플루메퀸(flumequine), 프룰리플록사신(prulifloxacin), 제미플록사신(gemifloxacin), 시프로플록사신(ciprofloxacin), 시타플록사신(sitafloxacin) 또는 클리나플록사신(clinafloxacin) 등일 수 있으나, 이에 제한되는 것은 아니다. 또한, 상기 아미노글라이코사이드계 항생제는 스트렙토마이신(streptomycin), 네오마이신(neomycin), 프라마이세틴(framycetin), 겐타마이신(gentamycin), 노보비오신(novobiocin), 가나마이신(kanamycin), 아미카신(amikacin), 시소마이신(sisomycin) 또는 스펙티노마이신(spectinomycin) 등일 수 있으나, 이에 제한되는 것은 아니다. In the present invention, the "antibiotic" may be rifampin, isoniazid, ethambutol, pyrazinamide (PZA), quinolone, or aminoglycoside, but is not limited thereto. Here, the quinolone antibiotic is nalidixic acid, marbofloxacin, oxolinic acid, moxifloxacin, trovafloxacin, gatifloxacin, Flumequine, prulifloxacin, gemifloxacin, ciprofloxacin, sitafloxacin, or clinafloxacin, etc., but may not be limited thereto. In addition, the aminoglycoside antibiotics include streptomycin, neomycin, framycetin, gentamycin, novobiocin, kanamycin, and amica. It may be syn (amikacin), sisomycin (sisomycin) or spectinomycin (spectinomycin), but is not limited thereto.
본 발명의 방법을 이용하는 경우 이용하여 비결핵 항산균에 의한 감염 여부 또는 감염 가능성을 진단할 수 있고, 질환의 경과, 예후 또는 치료 효과에 대하여도 추적할 수 있다.When the method of the present invention is used, it is possible to diagnose whether or not infection is caused by non-tuberculous mycobacteria, and the course of the disease, prognosis, or therapeutic effect can also be tracked.
본 발명의 정보 제공 방법에서 비결핵 항산균의 감염 질환에 관한 정의는 상기 본 발명의 바이오마커 조성물에 기재된 바와 중복되어 명세서의 과도한 혼잡을 피하기 위해 이하 그 기재를 생략한다.In the information providing method of the present invention, the definition of non-tuberculous mycobacterium infectious disease overlaps with that described in the biomarker composition of the present invention, and thus description thereof will be omitted below to avoid excessive congestion of the specification.
본 발명에서는 목적하는 개체의 생물학적 시료에 대하여 혈액 대사체의 발현 수준을 측정함으로써 비결핵 항산균에 의한 감염 질환의 발병 유무 또는 발병 가능성을 간단하고 용이하면서도 정확하게 예측할 수 있다. In the present invention, by measuring the expression level of a blood metabolite with respect to a biological sample of a target individual, the occurrence or possibility of an infectious disease caused by a non-tuberculous mycobacterium can be simply, easily and accurately predicted.
도 1은 본 발명의 일 실시예에서 항생제 치료 전의 마이코박테리움 아비움 복합체(MAC) 감염 폐 질환 환자와 건강한 사람의 혈청 시료에서 L-알기닌(L-Arginine)의 발현 수준을 비교한 그래프를 나타낸 것이다.1 is a graph comparing the expression level of L-arginine (L-Arginine) in serum samples of Mycobacterium avium complex (MAC)-infected lung disease patients and healthy people before antibiotic treatment in one embodiment of the present invention. it has been shown
도 2는 본 발명의 일 실시예에서 항생제 치료 전의 마이코박테리움 아비움 복합체(MAC) 감염 폐 질환 환자와 건강한 사람의 혈청 시료에서 알란토인(Allantoin)의 발현 수준을 비교한 그래프를 나타낸 것이다.Figure 2 shows a graph comparing the expression level of allantoin (Allantoin) in serum samples of patients with Mycobacterium avium complex (MAC)-infected lung disease and healthy people before antibiotic treatment in an embodiment of the present invention.
도 3은 본 발명의 일 실시예에서 항생제 치료 전의 마이코박테리움 아비움 복합체(MAC) 감염 폐 질환 환자와 건강한 사람의 혈청 시료에서 L-프롤린(L-Proline)의 발현 수준을 비교한 그래프를 나타낸 것이다.3 is a graph comparing the expression level of L-proline (L-Proline) in serum samples of patients with Mycobacterium avium complex (MAC)-infected lung disease and healthy people before antibiotic treatment in one embodiment of the present invention. it has been shown
도 4는 본 발명의 일 실시예에서 항생제 치료 전의 마이코박테리움 아비움 복합체(MAC) 감염 폐 질환 환자와 건강한 사람의 혈청 시료에서 L-트레오닌(L-Threonine)의 발현 수준을 비교한 그래프를 나타낸 것이다.Figure 4 is a graph comparing the expression level of L-threonine (L-Threonine) in serum samples of patients with Mycobacterium avium complex (MAC)-infected lung disease and healthy people before antibiotic treatment in an embodiment of the present invention. it has been shown
도 5는 본 발명의 일 실시예에서 항생제 치료 전의 마이코박테리움 아비움 복합체(MAC) 감염 폐 질환 환자와 건강한 사람의 혈청 시료에서 L-류신(L-Leucine)의 발현 수준을 비교한 그래프를 나타낸 것이다.5 is a graph comparing the expression level of L-leucine in serum samples of Mycobacterium avium complex (MAC)-infected lung disease patients and healthy people before antibiotic treatment in one embodiment of the present invention. it has been shown
도 6은 본 발명의 일 실시예에서 항생제 치료 전의 마이코박테리움 아비움 복합체(MAC) 감염 폐 질환 환자와 건강한 사람의 혈청 시료에서 L-라이신(L-Lycine)의 발현 수준을 비교한 그래프를 나타낸 것이다.6 is a graph comparing the expression level of L-lysine (L-Lycine) in serum samples of patients with Mycobacterium avium complex (MAC)-infected lung disease and healthy people before antibiotic treatment in one embodiment of the present invention. it has been shown
도 7은 본 발명의 일 실시예에서 항생제 치료 전의 마이코박테리움 아비움 복합체(MAC) 감염 폐 질환 환자와 건강한 사람의 혈청 시료에서 L-글루타메이트(L-Glutamate)의 발현 수준을 비교한 그래프를 나타낸 것이다.7 is a graph comparing the expression level of L-glutamate (L-Glutamate) in serum samples of patients with Mycobacterium avium complex (MAC)-infected lung disease and healthy people before antibiotic treatment in one embodiment of the present invention. it has been shown
도 8은 본 발명의 일 실시예에서 항생제 치료 전의 마이코박테리움 아비움 복합체(MAC) 감염 폐 질환 환자와 건강한 사람의 혈청 시료에서 L-트립토판(L-Tryptophan)의 발현 수준을 비교한 그래프를 나타낸 것이다.8 is a graph comparing the expression level of L-tryptophan (L-Tryptophan) in serum samples of patients with Mycobacterium avium complex (MAC)-infected lung disease and healthy people before antibiotic treatment in an embodiment of the present invention. it has been shown
도 9는 본 발명의 일 실시예에서 항생제 치료 전의 마이코박테리움 아비움 복합체(MAC) 감염 폐 질환 환자와 건강한 사람의 혈청 시료에서 N,N-디메틸글라이신(N,N-Dimethylglycine)의 발현 수준을 비교한 그래프를 나타낸 것이다.Figure 9 shows the expression level of N,N-dimethylglycine (N,N-Dimethylglycine) in serum samples from Mycobacterium avium complex (MAC)-infected lung disease patients and healthy people before antibiotic treatment in an embodiment of the present invention; A graph comparing them is shown.
도 10은 본 발명의 일 실시예에서 항생제 치료 전의 마이코박테리움 아비움 복합체(MAC) 감염 폐 질환 환자와 건강한 사람의 혈청 시료에서 호모세린(Homoserine)의 발현 수준을 비교한 그래프를 나타낸 것이다.10 shows a graph comparing the expression level of Homoserine in serum samples of Mycobacterium avium complex (MAC)-infected lung disease patients and healthy people before antibiotic treatment in an embodiment of the present invention.
도 11은 본 발명의 일 실시예에서 항생제 치료 전의 마이코박테리움 아비움 복합체(MAC) 감염 폐 질환 환자와 건강한 사람의 혈청 시료에서 D-2-하이드록시글루타릭산(D-2-hydroxyglutaric acid)의 발현 수준을 비교한 그래프를 나타낸 것이다.11 shows D-2-hydroxyglutaric acid in serum samples from patients with Mycobacterium avium complex (MAC)-infected lung disease and healthy people before antibiotic treatment in one embodiment of the present invention; ) shows a graph comparing the expression level.
도 12는 본 발명의 일 실시예에서 항생제 치료 전의 마이코박테리움 아비움 복합체(MAC) 감염 폐 질환 환자와 건강한 사람의 혈청 시료에서 L-아스파라긴(L-Asparagine)의 발현 수준을 비교한 그래프를 나타낸 것이다.12 is a graph comparing the expression level of L-asparagine (L-Asparagine) in serum samples of patients with Mycobacterium avium complex (MAC)-infected lung disease and healthy people before antibiotic treatment in one embodiment of the present invention. it has been shown
도 13은 본 발명의 일 실시예에서 항생제 치료 전의 마이코박테리움 아비움 복합체(MAC) 감염 폐 질환 환자와 건강한 사람의 혈청 시료에서 L-메티오닌(L-Methionine)의 발현 수준을 비교한 그래프를 나타낸 것이다.13 is a graph comparing the expression level of L-methionine (L-Methionine) in serum samples of patients with Mycobacterium avium complex (MAC)-infected lung disease and healthy people before antibiotic treatment in one embodiment of the present invention. it has been shown
도 14는 본 발명의 일 실시예에서 항생제 치료 전의 마이코박테리움 아비움 복합체(MAC) 감염 폐 질환 환자와 건강한 사람의 혈청 시료에서 콜린(Choline)의 발현 수준을 비교한 그래프를 나타낸 것이다.14 shows a graph comparing the expression level of choline in serum samples of patients with Mycobacterium avium complex (MAC)-infected lung disease and healthy people before antibiotic treatment in an embodiment of the present invention.
도 15는 본 발명의 일 실시예에서 항생제 치료 전의 마이코박테리움 아비움 복합체(MAC) 감염 폐 질환 환자와 건강한 사람의 혈청 시료에서 하이포잔틴(Hypoxanthine)의 발현 수준을 비교한 그래프를 나타낸 것이다.15 shows a graph comparing the expression level of hypoxanthine in serum samples of Mycobacterium avium complex (MAC)-infected lung disease patients and healthy people before antibiotic treatment in an embodiment of the present invention.
도 16은 본 발명의 일 실시예에서 항생제 치료 전의 마이코박테리움 아비움 복합체(MAC) 감염 폐 질환 환자와 건강한 사람의 혈청 시료에서 S-락테이트(S-Lactate)의 발현 수준을 비교한 그래프를 나타낸 것이다.16 is a graph comparing the expression level of S-lactate (S-Lactate) in serum samples of patients with Mycobacterium avium complex (MAC)-infected lung disease and healthy people before antibiotic treatment in an embodiment of the present invention; is shown.
도 17은 본 발명의 일 실시예에서 항생제 치료 전의 마이코박테리움 아비움 복합체(MAC) 감염 폐 질환 환자와 건강한 사람의 혈청 시료에서 L-말산(L-Malic acid)의 발현 수준을 비교한 그래프를 나타낸 것이다.17 is a graph comparing the expression level of L-malic acid in serum samples of patients with Mycobacterium avium complex (MAC)-infected lung disease and healthy people before antibiotic treatment in one embodiment of the present invention; is shown.
도 18은 본 발명의 일 실시예에서 항생제 치료 전의 마이코박테리움 아비움 복합체(MAC) 감염 폐 질환 환자와 건강한 사람의 혈청 시료에서 글리세롤 3-포스페이트(Glycerol 3-phosphate)의 발현 수준을 비교한 그래프를 나타낸 것이다.18 is a comparison of the expression level of glycerol 3-phosphate in serum samples of mycobacterium avium complex (MAC)-infected lung disease patients and healthy people before antibiotic treatment in one embodiment of the present invention. graph is shown.
도 19는 본 발명의 일 실시예에서 항생제 치료 전의 마이코박테리움 아비움 복합체(MAC) 감염 폐 질환 환자와 건강한 사람의 혈청 시료에서 타우린(Taurine)의 발현 수준을 비교한 그래프를 나타낸 것이다.19 shows a graph comparing the expression level of taurine in serum samples of patients with Mycobacterium avium complex (MAC)-infected lung disease and healthy people before antibiotic treatment in an embodiment of the present invention.
도 20은 본 발명의 일 실시예에서 항생제 치료 전의 마이코박테리움 아비움 복합체(MAC) 감염 폐 질환 환자와 건강한 사람의 혈청 시료에서 L-세린(L-Serine)의 발현 수준을 비교한 그래프를 나타낸 것이다.20 is a graph comparing the expression level of L-serine (L-Serine) in serum samples from patients with Mycobacterium avium complex (MAC)-infected lung disease and healthy people before antibiotic treatment in one embodiment of the present invention. it has been shown
본 발명의 일 구현 예에 따르면, 목적하는 개체로부터 분리된 생물학적 시료에서 대사체의 발현 수준을 측정하는 단계를 포함하는 비결핵 항산균의 감염 또는 감염 질환을 진단하기 위한 정보 제공 방법에 관한 것이다. According to one embodiment of the present invention, it relates to a method for providing information for diagnosing infection or infectious disease of non-tuberculous mycobacteria, comprising measuring the expression level of a metabolite in a biological sample isolated from a target subject.
본 발명에서 상기 대사체는 혈액, 바람직하게는 혈청 기원의 액상 시료로부터 수득한 대사물질인 것이 바람직하고, 구체적인 예를 들면, 아미노산(amino acid), 아미노산 유도체, 알란토인(Allantoin), N,N-디메틸글라이신(N,N-Dimethylglycine), 2-하이드록시글루타릭산(2-hydroxyglutaric acid), 콜린(Choline), 하이포잔틴(Hypoxanthine), 락테이트(Lactate), 말산(Malic acid) 및 글리세롤 3-포스페이트(Glycerol 3-phosphate)로 이루어진 군에서 선택된 1종 이상을 포함할 수 있다. 여기서 상기 아미노산 및 그 유도체는 알기닌(Arginine), 프롤린(Proline), 트레오닌(Threonine), 류신(Leucine), 라이신(Lycine), 글루타메이트(Glutamate), 트립토판(Tryptophan), 아스파라긴(Asparagine), 메티오닌(Methionine), 세린(Serine), 호모세린(Homoserine) 및 타우린(Taurine)으로 이루어진 군에서 선택된 1종 이상을 포함할 수 있다. 또한, 본 발명에서 상기 2-하이드록시글루타릭산(2-hydroxyglutaric acid)은 D-형태(D-form)이고, 상기 락테이트(Lactate)는 S-형태(S-form)이며, 상기 말산(Malic acid)은 L-형태(L-form)이고, 상기 아미노산은 (L)-형태((L)-form)일 수 있다. In the present invention, the metabolite is preferably a metabolite obtained from a liquid sample derived from blood, preferably serum, and specific examples include amino acids, amino acid derivatives, allantoin, N,N- Dimethylglycine (N,N-Dimethylglycine), 2-hydroxyglutaric acid, Choline, Hypoxanthine, Lactate, Malic acid and Glycerol 3- It may include one or more selected from the group consisting of phosphate (Glycerol 3-phosphate). Here, the amino acid and its derivatives are Arginine, Proline, Threonine, Leucine, Lysine, Glutamate, Tryptophan, Asparagine, Methionine ), serine (Serine), homoserine (Homoserine) and taurine (Taurine) may include one or more selected from the group consisting of. In addition, in the present invention, the 2-hydroxyglutaric acid is D-form, the lactate is S-form, and the malic acid ( Malic acid) may be in the L-form (L-form), and the amino acid may be in the (L)-form ((L)-form).
이하, 본 발명을 하기의 실시예에 의해 상세히 설명한다. 단, 하기 실시예는 본 발명을 예시하는 것일 뿐, 본 발명의 내용이 하기 실시예에 의해 한정되는 것은 아니다.Hereinafter, the present invention will be described in detail by the following examples. However, the following examples are only illustrative of the present invention, and the content of the present invention is not limited by the following examples.
실시예Example
[실험예 1] MAC 감염 환자와 건강한 사람의 시료 수집[Experimental Example 1] Collecting samples from MAC-infected patients and healthy people
2012년 1월부터 2016년 8월까지 기간 동안 대략 6년간 서울 삼성병원에서 수집한 마이코박테리움 아비움 복합체(Mycobacterium avium complex) (avium : 46명, intracellulare : 50명, 총 96명) 감염 폐 질환 환자의 항생제 치료 전의 혈청 샘플 96개와 건강한 사람의 혈청 30개를 준비하였다. 단, 상기 마이코박테리움 아비움 복합체 감염 폐 질환 환자 96명 중 기관지 확장증형 (nodular bronchiectatic form) 환자는 66명이고, 상엽 공동형(upper lobe cavitary form) 환자는 30명이었다. Mycobacterium avium complex ( avium : 46, intracellulare : 50, total 96) infected lungs collected at Samsung Hospital in Seoul for approximately 6 years from January 2012 to August 2016 96 sera samples from diseased patients before antibiotic treatment and 30 sera from healthy people were prepared. However, among the 96 patients with the Mycobacterium avium complex-infected lung disease, 66 patients had nodular bronchiectatic form and 30 patients had upper lobe cavitary form.
[실험예 2] 시료에 대한 전처리[Experimental Example 2] Pretreatment of samples
먼저, 상기 실험예 1에서 얻어진 혈청 시료 (50 μl)에 300 μl 클로로포름, 150 μl 메탄올 (chloroform-methanol, 2:1, v/v, 4 ℃)을 첨가하고 30초 동안 섞어 주었다. 여기에 150 μl 물을 첨가하고 30 초 동안 섞은 뒤 ICE에 넣어 10 분간 방치하여 추출하였다. 이후, 원심분리기기를 이용하여 10 분간 13,000 rpm, 4 ℃에서 원심분리한 뒤 상층액(250 μl) 분리해내어 Speed vacuum (full vacuum, no temp, 5hours)을 이용하여 건조하여 이하의 대사체 분석 전까지 -20 ℃에서 보관하였다. 질량 분석기 분석을 위해 건조된 시료를 250 μl 아세토니트릴-H2O(Acetonitrile-H2O)(75:25, v/v)에 재용해 후, 존재할 가능성이 있는 불순물 제거를 위하여 필터 튜브(Filter tube)(Costar 8169)를 이용하여 여과한 후 분석을 진행하였다. 기계 품질 관리(Machinery Quality Control; MQC)로, MS/MS 기기상태를 체크하기 위하여 혈청 샘플과 같은 전 처리방법으로 건강한 사람의 혈청을 기계 품질 관리(MQC)의 샘플로 사용하여 배치 당 4회 반복 분석하였다. 시료 품질 관리(Sample Quality Control; SQC)를 위하여 각 배치 안에서 시료 간의 차이를 비교하기 위해 시료 당 20 μl씩 모아 시료 품질 관리를 제작하여 배치 당 4회 반복 분석하였다.First, 300 μl chloroform and 150 μl methanol (chloroform-methanol, 2:1, v/v, 4 ℃) were added to the serum sample (50 μl) obtained in Experimental Example 1 and mixed for 30 seconds. 150 μl of water was added here, mixed for 30 seconds, put in ICE, and left for 10 minutes for extraction. Then, after centrifugation at 13,000 rpm, 4 °C for 10 minutes using a centrifugal separator, the supernatant (250 μl) was separated and dried using a speed vacuum (full vacuum, no temp, 5hours) to analyze the following metabolites It was stored at -20 °C until For mass spectrometry analysis, re-dissolve the dried sample in 250 μl acetonitrile-H 2 O (75:25, v/v), and filter tube to remove any impurities that may be present. (Costar 8169) was used for filtration and analysis was performed. With Machinery Quality Control (MQC), it is a pre-processing method such as a serum sample to check the condition of MS/MS equipment. Using serum from a healthy person as a sample for machine quality control (MQC), repeat 4 times per batch analyzed. For sample quality control (SQC), 20 μl per sample was collected to compare differences between samples in each batch, and sample quality control was prepared and analyzed 4 times per batch.
[실험예 3][Experimental Example 3] HPLC-Triple Quad-MS를 통한 대사체 분석Metabolite analysis by HPLC-Triple Quad-MS
혈청에서 처리한 분석 시료 내의 극성 대사체를 분석하기 위하여 크로마토그래피-텐덤 질량분석기(HPLC-MS/MS)를 이용하여 분석을 진행하였다. 사용된 장비는 Agilent 1200 HPLC와 Sciex API4000 triple quadrupole MS를 이용하였다. 친수성 상호 작용을 위한 크로마토그래피 조건으로는 Luna PFPP(2.0 x 150 mm, 3μm, Phenomenex) 컬럼을 이용하여 20 ℃에서 용매에 따른 2가지 방법으로 기울기 용리를 이용하여 극성 대사체들을 분리하였다. 첫 번째 이동상으로는 (A) H2O (v/v) 및 (B) 아세토니트릴 (v/v)을 이용하였고, 두 번째 이동상으로는 (A) H2O (v/v, 0.1% formic acid) 및 (B) 아세토니트릴 (v/v)을 이용하였으며, 각각 조건의 기울기 용리는 총 분석 시간을 15분으로 하여 아래 표 1과 동일하게 수행하였다. 분무기 가스(Ion-Source Gas 1/2) 단위는 50/50 임의 단위(arbitrary unit)이었으며, 가스 커튼(Curtain Gas)의 단위는 25 임의 단위(arbitrary unit)이었다. 소스 온도(Source temperature)는 500 ℃이고, 이온 스프레이 부유 전압(Ion-spray Floating Voltage)은 5,5 kV(negative -4.5kV)이며, 매스 범위(Mass range)는 50 ~ 1000 m/z이었다. 시료 주입은 HTC_PAL system/CTC analytics auto-sampler를 이용하여 3 μl씩 주입하였으며, 텐덤 질량 분석기 조건 (예약 다중 반응 검지법; Scheduled Multiple Reaction Monitoring, sMRM)은 아래 표 2 내지 5와 같이 수행하였다. 단, 하기 표 2 내지 5에서 m/z는 질량 대 전하비(mass to charge ratio)를 의미하고, RT는 머무름 시간(Retention time)을 의미하며, CE는 충돌 에너지(Collision energy)를 의미하고, (+)는 양이온 모드를, (-)는 음이온 모드를 의미하며, sMRM 분석을 통해 얻어진 결과는 Sciex의 정량 분석 소프트웨어(Quantitative Analysis Software)를 통하여 로우 데이터(raw data)를 계산하였고, MQC data 평균값을 이용하여 상대 표준 편차(RSD<20)이하의 대사체를 산출 하였다.In order to analyze the polar metabolites in the analysis sample treated with serum, the analysis was performed using chromatography-tandem mass spectrometry (HPLC-MS/MS). The equipment used was an Agilent 1200 HPLC and a Sciex API4000 triple quadrupole MS. As chromatography conditions for the hydrophilic interaction, polar metabolites were separated by gradient elution in two methods depending on the solvent at 20 °C using a Luna PFPP (2.0 x 150 mm, 3 μm, Phenomenex) column. (A) H 2 O (v/v) and (B) acetonitrile (v/v) were used as the first mobile phase, and (A) H 2 O (v/v, 0.1% formic acid) as the second mobile phase and (B) acetonitrile (v/v) were used, and the gradient elution of each condition was performed in the same manner as in Table 1 below with a total analysis time of 15 minutes. The unit of the Ion-Source Gas 1/2 was 50/50 arbitrary units, and the unit of the Curtain Gas was 25 arbitrary units. The source temperature was 500 °C, the ion-spray floating voltage was 5.5 kV (negative -4.5 kV), and the mass range was 50 ~ 1000 m/z. Samples were injected by 3 μl using HTC_PAL system/CTC analytics auto-sampler, and tandem mass spectrometry conditions (Scheduled Multiple Reaction Monitoring, sMRM) were performed as shown in Tables 2 to 5 below. However, in Tables 2 to 5, m/z means mass to charge ratio, RT means retention time, CE means collision energy, (+) means positive ion mode and (-) means negative ion mode. For the results obtained through sMRM analysis, raw data was calculated through Sciex's Quantitative Analysis Software, and MQC data average value was used to calculate metabolites with a relative standard deviation (RSD<20) or less.
시간(분)hours (minutes) 이동상 A(%)Mobile phase A (%) 이동상 B(%)Mobile phase B (%) 유속(mL/min)Flow rate (mL/min)
00 100100 00 0.350.35
88 7373 2727 0.350.35
99 1515 8585 0.350.35
1010 100100 00 0.350.35
1515 100100 00 0.350.35
물 방법 (+)(Water method (+)), 대사체 21종 Water method (+), 21 metabolites
대사체 종류(Compounds)Metabolites (Compounds) m/zm/z Product ionproduct ion RTRT CECE
타이로신(L-Tyrosine)Tyrosine (L-Tyrosine) 182182 7777 2.82.8 4141
시스타티오닌(L-Cystathionine)Cystathionine (L-Cystathionine) 223223 134134 0.90.9 1111
베타인(Betaine)Betaine 118118 5858 1.321.32 3939
티아민(Thymine)Thymine 127127 110110 5.435.43 2121
오르니틴(Ornithine)Ornithine 133133 7070 0.950.95 2525
구아닌(Guanine)Guanine 152152 110110 2.22.2 2727
히스티딘(Histidine)Histidine 156156 110110 1.321.32 1212
아세틸오르니틴(N-Acetylornithine)Acetylornithine (N-Acetylornithine) 175175 115115 1.141.14 1414
글루코사민(Glucosamine)Glucosamine 180180 162162 1.41.4 1010
우라실(Uracil)Uracil 113113 7070 1.851.85 2323
디옥시우리딘(Deoxyuridine)Deoxyuridine 229229 113113 6.216.21 1111
씨티딘(Cytidine)Cytidine 244244 112112 2.442.44 1212
디옥시아데노신(Deoxyadenosine)Deoxyadenosine 252252 136136 8.58.5 2020
디옥시이노신(Deoxyinosine)Deoxyinosine 253253 137137 6.586.58 1212
아데노신(Adenosine)Adenosine 268268 136136 7.727.72 2727
디옥시구아노신(Deoxyguanosine)Deoxyguanosine 268268 152152 6.846.84 1515
이노신(Inosine)Inosine 269269 137137 6.246.24 1414
구아노신(Guanosine)Guanosine 284284 152152 6.556.55 1717
잔토신(Xanthosine)Xanthosine 285285 153153 6.976.97 2020
싸이클릭 AMP(Cyclic AMP)Cyclic AMP 328328 287287 0.860.86 99
SAHSAH 385385 136136 0.460.46 1919
물 방법 (-)(Water method (-)), 대사체 12종 Water method (-)), 12 metabolites
대사체 종류(Compounds)Metabolites (Compounds) m/zm/z Product ionproduct ion RTRT CECE
글루코스(D-Glucose)Glucose (D-Glucose) 179179 8989 0.920.92 -12-12
하이드록시뷰티레이트(3-hydroxybutyric acid)Hydroxybutyrate (3-hydroxybutyric acid) 103103 4141 0.960.96 -32-32
타우린(Taurine)Taurine 124124 8080 0.90.9 -16-16
안트라닐레이트(Anthranilate)Anthranilate 136136 9292 3.683.68 -16-16
하이드록시벤조에이트(p-Hydroxybenzoate)Hydroxybenzoate (p-Hydroxybenzoate) 137137 9393 1.61.6 -21-21
시투를린(Citrulline)Citrulline 174174 131131 1.211.21 -13-13
하이드록시페닐파이루베이트(p-Hydroxybenzoate)Hydroxyphenylpyruvate (p-Hydroxybenzoate) 179179 107107 0.970.97 -11-11
마이오이노시톨(myo-Inositol)myo-Inositol 179179 161161 0.940.94 -15-15
티미딘(Thymidine)Thymidine 241241 125125 7.027.02 -10-10
우리딘(Uridine)Uridine 243243 111111 4.224.22 -12-12
니코티네이트(Nicotinate)Nicotinate 122122 7878 1.541.54 -14-14
우레이트(Uric acid)Uric acid 167167 124124 1.351.35 -22-22
포름산 방법 (+)(Formic acid method (+)), 대사체 32종Formic acid method (+), 32 metabolites
대사체 종류(Compounds)Metabolites (Compounds) m/zm/z Product ionproduct ion RTRT CECE
세린(L-Serine)Serine (L-Serine) 106106 6060 0.90.9 1515
프롤린(L-Proline)Proline (L-Proline) 116116 7070 1.131.13 2121
발린(L-Valine)L-Valine 118118 7272 1.411.41 1515
트레오닌(L-Threonine)Threonine (L-Threonine) 120120 7474 0.960.96 1515
이소류신(L-isoLeucine)Isoleucine (L-isoLeucine) 132132 8686 1.931.93 1515
류신(L-Leucine)Leucine (L-Leucine) 132132 8686 2.32.3 1515
아스파라긴(L-Asparagine)Asparagine (L-Asparagine) 133133 7474 0.90.9 1717
글루타민(L-Glutamine)Glutamine (L-Glutamine) 147147 8484 0.950.95 2323
라이신(L-Lycine)Lysine (L-Lycine) 147147 8484 0.740.74 2323
글루타메이트(L-Glutamate)Glutamate (L-Glutamate) 148148 8484 1One 2323
메티오닌(L-Methionine)Methionine (L-Methionine) 150150 104104 1.81.8 1111
페닐알라닌(L-Phenylalanine)Phenylalanine (L-Phenylalanine) 166166 120120 6.416.41 1717
알지닌(L-Arginine)Arginine (L-Arginine) 175175 7070 0.80.8 3535
트립토판(L-Tryptophan)Tryptophan (L-Tryptophan) 205205 188188 8.48.4 1313
다이메틸글라이신(N,N-Dimethylglycine)Dimethylglycine (N,N-Dimethylglycine) 104104 5858 1.211.21 2727
콜린(Choline)Choline 104104 6060 0.90.9 1919
글라이신(Glycine)Glycine 7676 3030 1.061.06 1616
폴레이트(Folate)Folate 442442 295295 9.589.58 1919
아데닌(Adenine)Adenine 136136 119119 1.751.75 2424
호모시스테인(Homocysteine)Homocysteine 136136 9090 1.261.26 1515
하이포잔틴(Hypoxanthine)Hypoxanthine 137137 110110 2.82.8 2929
잔틴(Xanthine)Xanthine 153153 110110 2.52.5 2323
알란토인(Allantoin)Allantoin 159159 9999 1.171.17 1313
사이토신(Cytosine)Cytosine 112112 9595 0.980.98 1717
호모세린(Homoserine)Homoserine 120120 5656 1.031.03 2727
티아민(Thiamine)Thiamine 265265 122122 0.960.96 1717
시스테인(Cysteine)Cysteine 122122 5959 1.241.24 2727
CMPCMP 324324 112112 1.681.68 1616
UMPUMP 325325 9797 33 4949
AMPAMP 348348 136136 1.991.99 2121
IMPIMP 349349 137137 5.75.7 1717
스펄민(Spermine)Spermine 203203 112112 0.530.53 2727
포름산 방법 (-)(Formic acid method (-)), 대사체 24종Formic acid method (-), 24 metabolites
대사체 종류(Compounds)Metabolites (Compounds) m/zm/z Product ionproduct ion RTRT CECE
아스팔테이트(L-Aspartate)Aspartate (L-Aspartate) 132132 8888 0.960.96 -17-17
락테이트((S)-Lactate))Lactate ((S)-Lactate)) 8989 4343 1.741.74 -18-18
포스포글리세라이트(3-Phosphoglycerate)Phosphoglycerate (3-Phosphoglycerate) 185185 9797 2.112.11 -22-22
석시네이트(Succinate)Succinate 117117 7373 3.883.88 -18-18
말레이트(L-Malic acid)Malate (L-Malic acid) 133133 115115 1.781.78 -16-16
시트레이트(Citrate)Citrate 191191 111111 3.583.58 -12-12
하이드로글루타레이트(D-2-Hydroyglutaric acid)Hydroglutarate (D-2-Hydroyglutaric acid) 147147 129129 1.031.03 -14-14
GTPGTP 522522 424424 1.61.6 -30-30
아세틸포스페이트(Acetylphosphate)Acetylphosphate 139139 7979 1.651.65 -22-22
칼바모일포스페이트(Carbamoyl-phosphate)Carbamoyl-phosphate 140140 7979 0.90.9 -22-22
글리세라이트(Glycerate) Glycerate 105105 7575 1.241.24 -15-15
포스포에놀파이루베이트(Phosphoenolpyruvate)Phosphoenolpyruvate 167167 7979 2.32.3 -16-16
디하이드록시아세톤포스페이트(Dihydroxyacetone phosphate)Dihydroxyacetone phosphate 169169 7979 1.71.7 -38-38
글리세롤 3-포스페이트(Glycerol 3-Phosphate)Glycerol 3-Phosphate 171171 7979 1.51.5 -22-22
시키메이트(Shikimate)Shikimate 173173 9393 1.651.65 -16-16
알란토에이트(Allontoate)Allontoate 175175 132132 1.051.05 -12-12
디옥시리보스 1-포스페이트(Deoxyrebose 1-Phosphate)Deoxyrebose 1-Phosphate 213213 7979 1.61.6 -33-33
리불로스 5-포스페이트(D-Ribulose 5-Phosphate)D-Ribulose 5-Phosphate 229229 7979 1.31.3 -48-48
글루코스 6-포스페이트(Glucose 6-Phosphate)Glucose 6-Phosphate 259259 7979 1.731.73 -40-40
프룩토스 1,6-비스포스페이트(Fructose 1,6-Bisphosphate)Fructose 1,6-Bisphosphate 339339 271271 0.980.98 -18-18
dGMPdGMP 346346 7979 2.022.02 -20-20
PRPPPRPP 389389 291291 1.41.4 -18-18
이타코네이트(Itaconate)Itaconate 129129 8585 6.46.4 -14-14
프룩토스 6-포스페이트(Fructose 6-Phosphate)Fructose 6-Phosphate 259259 7979 1.231.23 -54-54
[실험예 4] 비결핵 항산균 감염자의 혈청 시료 내 대사체 분석 결과[Experimental Example 4] Metabolite analysis results in serum samples of non-tuberculous mycobacteria infection
항생제 치료 전의 마이코박테리움 아비움 복합체(MAC) 감염 폐 질환 환자와 건강한 사람의 대사체 농도를 비교하기 위해 다음의 통계 검정 2가지 방법으로 Metaboanalyst(data 통계사이트)와 SPSS 통계 프로그램을 이용하여 데이터를 산출하였고, 그 결과를 이용하여 치료 전의 마이코박테리움 아비움 복합체(MAC) 감염 폐 질환 환자와 건강한 사람을 구분 할 수 있는 질병 관련 대사체 총 20 종(p-value>0.05)을 각각의 p-value와 건강한 사람 대비 발현 수준의 배수 변화(Fold change) 값을 토대로 선정하여 그 결과를 하기 표 6 및 도 1 내지 20에 나타내었다. 단, 도 1 내지 20에서, HC는 건강한 사람 30명의 혈청 샘플에서 각 대사체의 발현 수준을 나타낸 것이고, Dx0=Tx0(Success & Fail), 항생제 치료 시작전의 마이코박테리움 아비움 복합체(MAC) 감염 환자 96명의 혈청 샘플에서 각 대사체의 발현 수준을 나타낸 것이다. 또한, 유의성 unpaired t-test에서 *P<0.05; **P<0.01; ***P<0.001를 의미한다.To compare metabolite concentrations in patients with Mycobacterium avium complex (MAC)-infected lung disease and healthy people before antibiotic treatment, the following statistical tests were performed using Metaboanalyst (data statistics site) and SPSS statistical program. was calculated, and using the results, a total of 20 disease-related metabolites (p-value >0.05) that can distinguish patients with mycobacterium avium complex (MAC)-infected lung disease before treatment and healthy people were obtained for each It was selected based on the p-value and the fold change value of the expression level compared to a healthy person, and the results are shown in Table 6 and FIGS. 1 to 20 below. However, in FIGS. 1 to 20, HC represents the expression level of each metabolite in serum samples of 30 healthy people, Dx0 = Tx0 (Success & Fail), Mycobacterium avium complex (MAC) before the start of antibiotic treatment Expression levels of each metabolite in serum samples from 96 infected patients are shown. In addition, *P<0.05 in the significance unpaired t-test; **P<0.01; *** means P<0.001.
건강한 사람 대비 치료전 MAC 감염 환자에서 증가된 대사체Increased metabolites in patients with pre-treatment MAC infection compared to healthy individuals
대사체 종류(Compounds)Metabolites (Compounds) 유의성 (p-value)Significance (p-value) Fold ChangeFold Change
알기닌(L-Arginine)Arginine (L-Arginine) 0.01370.0137 1.361.36
알란토인(Allantoin)Allantoin 0.00610.0061 1.121.12
건강한 사람 대비 치료전 MAC 감염 환자에서 감소된 대사체Reduced metabolites in patients with pre-treatment MAC infection compared to healthy individuals
프롤린(L-Proline)Proline (L-Proline) 0.03110.0311 0.830.83
트레오닌(L-Threonine)Threonine (L-Threonine) 0.0010.001 0.710.71
류신(L-Leucine)Leucine (L-Leucine) 0.03990.0399 0.790.79
라이신(L-Lysine)Lysine (L-Lysine) 0.01090.0109 0.750.75
글루타메이트(L-Glutamate)Glutamate (L-Glutamate) 0.01240.0124 0.710.71
트립토판(L-Tryptophan)Tryptophan (L-Tryptophan) 0.00080.0008 0.780.78
N,N-다이메틸글라이신(N,N-Dimethylglycine)N,N-Dimethylglycine (N,N-Dimethylglycine) 0.00010.0001 0.720.72
호모세린(Homoserine)Homoserine 0.00070.0007 0.690.69
D-2-하이드록시글루타릭 엑시드(D-2-hydroxyglutaric acid)D-2-hydroxyglutaric acid 0.03160.0316 0.680.68
아스파라긴(L-Asparagine)Asparagine (L-Asparagine) 0.03390.0339 0.740.74
메티오닌(L-Methionine)Methionine (L-Methionine) 0.00080.0008 0.650.65
콜린(Choline)Choline 0.0010.001 0.730.73
하이포잔틴(Hypoxanthine)Hypoxanthine 0.00010.0001 0.360.36
(S)-락테이트(S)-Lactate)(S)-Lactate (S)-Lactate) 0.00010.0001 0.440.44
말릭 엑시드(L-Malic acid)Malic acid (L-Malic acid) 0.00170.0017 0.340.34
글리세롤 3-포스페이트(Glycerol 3-phosphate)Glycerol 3-phosphate 0.01380.0138 0.420.42
타우린(Taurine)Taurine 0.00010.0001 0.370.37
세린(L-Serine)Serine (L-Serine) 0.05510.0551 0.840.84
상기 표 6 및 도 1 내지 20에서 보는 바와 같이, 혈액 대사체 중 L-알기닌(L-Arginine) 및 알란토인(Allantoin)은 건강한 사람 대비 비결핵 항산균 감염자에서 유의적으로 그 발현이 증가하였고, 그 외에 L-프롤린(L-Proline), L-트레오닌(L-Threonine), L-류신(L-Leucine), L-라이신(L-Lycine), L-글루타메이트(L-Glutamate), L-트립토판(L-Tryptophan), N,N-디메틸글라이신(N,N-Dimethylglycine), 호모세린(Homoserine), D-2-하이드록시글루타릭산(D-2-hydroxyglutaric acid), L-아스파라긴(L-Asparagine), L-메티오닌(L-Methionine), 콜린(Choline), 하이포잔틴(Hypoxanthine), S-락테이트(S-Lactate), L-말산(L-Malic acid), 글리세롤 3-포스페이트(Glycerol 3-phosphate), 타우린(Taurine) 및 L-세린(L-Serine)은 건강한 사람 대비 비결핵 항산균 감염자에서 유의적으로 그 발현이 감소한 것을 확인할 수 있었다. As shown in Table 6 and FIGS. 1 to 20, the expression of L-arginine and allantoin among blood metabolites was significantly increased in non-tuberculous mycobacterium-infected patients compared to healthy people, and the In addition, L-Proline, L-Threonine, L-Leucine, L-Lycine, L-Glutamate, L-Tryptophan ( L-Tryptophan), N,N-Dimethylglycine, Homoserine, D-2-hydroxyglutaric acid, L-Asparagine ), L-methionine (L-Methionine), choline (Choline), hypoxanthine (Hypoxanthine), S-lactate (S-Lactate), L-malic acid (L-Malic acid), glycerol 3-phosphate (Glycerol 3- phosphate), taurine (Taurine) and L-serine (L-Serine) were confirmed that the expression was significantly reduced in the non-tuberculous mycobacterium-infected patient compared to healthy people.
이를 통하여 대사체로, L-알기닌(L-Arginine), 알란토인(Allantoin), L-프롤린(L-Proline), L-트레오닌(L-Threonine), L-류신(L-Leucine), L-라이신(L-Lycine), L-글루타메이트(L-Glutamate), L-트립토판(L-Tryptophan), N,N-디메틸글라이신(N,N-Dimethylglycine), 호모세린(Homoserine), D-2-하이드록시글루타릭산(D-2-hydroxyglutaric acid), L-아스파라긴(L-Asparagine), L-메티오닌(L-Methionine), 콜린(Choline), 하이포잔틴(Hypoxanthine), S-락테이트(S-Lactate), L-말산(L-Malic acid), 글리세롤 3-포스페이트(Glycerol 3-phosphate), 타우린(Taurine) 및 L-세린(L-Serine)을 비결핵 항산균에 의한 감염 또는 감염 질환을 진단하기 위한 바이오마커로 사용할 수 있음을 알 수 있었다.Through this, metabolites, L-Arginine, Allantoin, L-Proline, L-Threonine, L-Leucine, L-Lysine ( L-Lycine), L-Glutamate, L-Tryptophan, N,N-Dimethylglycine, Homoserine, D-2-hydroxyglycine Rutaric acid (D-2-hydroxyglutaric acid), L-Asparagine (L-Asparagine), L-Methionine (L-Methionine), Choline (Choline), Hypoxanthine (Hypoxanthine), S-Lactate (S-Lactate), L-Malic acid, glycerol 3-phosphate, taurine, and L-serine for diagnosing infections or infectious diseases caused by non-tuberculous mycobacteria It was found that it can be used as a marker.
본 발명은 비결핵 항산균에 의한 감염 또는 감염 질환을 진단하기 위한 바이오마커와, 상기 진단을 위한 키트 또는 진단을 위한 정보 제공 방법에 관한 것이다. The present invention relates to a biomarker for diagnosing infection or infectious disease caused by non-tuberculous mycobacteria, and a kit for the diagnosis or a method for providing information for diagnosis.

Claims (23)

  1. 아미노산(amino acid), 아미노산 유도체, 알란토인(Allantoin), N,N-디메틸글라이신(N,N-Dimethylglycine), 2-하이드록시글루타릭산(2-hydroxyglutaric acid), 콜린(Choline), 하이포잔틴(Hypoxanthine), 락테이트(Lactate), 말산(Malic acid) 및 글리세롤 3-포스페이트(Glycerol 3-phosphate)로 이루어진 군에서 선택된 1종 이상의 대사체를 포함하는, 비결핵 항산균의 감염 또는 감염 질환의 진단용 바이오마커 조성물.Amino acids, amino acid derivatives, allantoin, N,N-dimethylglycine, 2-hydroxyglutaric acid, choline, hypoxanthine ( Hypoxanthine), lactate (Lactate), malic acid (Malic acid) and glycerol 3-phosphate (Glycerol 3-phosphate) containing one or more metabolites selected from the group consisting of, non-tuberculous mycobacteria for the diagnosis of infection or infectious disease biomarker composition.
  2. 제1항에 있어서,According to claim 1,
    상기 아미노산 및 아미노산 유도체는 알기닌(Arginine), 프롤린(Proline), 트레오닌(Threonine), 류신(Leucine), 라이신(Lycine), 글루타메이트(Glutamate), 트립토판(Tryptophan), 아스파라긴(Asparagine), 메티오닌(Methionine), 세린(Serine), 호모세린(Homoserine) 및 타우린(Taurine)으로 이루어진 군에서 선택된 1종 이상을 포함하는, 비결핵 항산균의 감염 또는 감염 질환의 진단용 바이오마커 조성물.The amino acids and amino acid derivatives are Arginine, Proline, Threonine, Leucine, Lysine, Glutamate, Tryptophan, Asparagine, Methionine. , Serine (Serine), homoserine (Homoserine) and taurine (Taurine) comprising at least one selected from the group consisting of, non-tuberculous mycobacterium infection or biomarker composition for diagnosis of infectious diseases.
  3. 제1항에 있어서,According to claim 1,
    상기 아미노산은 L-형태(L-form)인, 비결핵 항산균의 감염 또는 감염 질환의 진단용 바이오마커 조성물.The amino acid is L-form (L-form), a biomarker composition for diagnosis of non-tuberculous mycobacteria infection or infectious disease.
  4. 제1항에 있어서, According to claim 1,
    상기 대사체는 목적하는 개체의 전혈(whole blood), 혈장(plasma) 또는 혈청(serum) 유래인 것인, 비결핵 항산균의 감염 또는 감염 질환의 진단용 바이오마커 조성물.The metabolite is a biomarker composition for diagnosis of non-tuberculous mycobacterium infection or infectious disease, which is derived from whole blood, plasma or serum of a target individual.
  5. 제1항에 있어서,According to claim 1,
    상기 비결핵 항산균은 마이코박테리움 아비움(M. avium), 마이코박테리움 압세수스(M. abscessus), 마이코박테리움 플라베센스(M. flavescence), 마이코박테리움 아프리카눔(M. africanum), 마이코박테리움 보비스(M. bovis), 마이코박테리움 첼로네(M. chelonae), 마이코박테리움 셀라툼(M. celatum), 마이코박테리움 포르투이툼(M. fortuitum), 마이코박테리움 고르도네(M. gordonae), 마이코박테리움 가스트리(M. gastri), 마이코박테리움 헤모필룸(M. haemophilum), 마이코박테리움 인트라셀루라레(M. intracellulare), 마이코박테리움 칸사시이(M. kansasii), 마이코박테리움 말모엔스(M. malmoense), 마이코박테리움 마리눔(M. marinum), 마이코박테리움 스줄가이(M. szulgai), 마이코박테리움 테레(M. terrae), 마이코박테리움 스크로풀라세움(M. scrofulaceum), 마이코박테리움 울서란스(M. ulcerans), 마이코박테리움 시미애(M. simiae) 및 마이코박테리움 제노피(M. xenopi)로 구성된 군으로부터 선택되는, 비결핵 항산균의 감염 또는 감염 질환의 진단용 바이오마커 조성물.The non-tuberculous mycobacteria are Mycobacterium avium (M. avium), Mycobacterium abscessus (M. abscessus), Mycobacterium flavescens (M. flavescence), Mycobacterium africanum (M. africanum), Mycobacterium bovis (M. bovis), Mycobacterium cellone (M. chelonae), Mycobacterium cellatum (M. celatum), Mycobacterium portuitum (M. fortuitum), Mycobacterium gordonae (M. gordonae), Mycobacterium gastri (M. gastri), Mycobacterium haemophilum (M. haemophilum), Mycobacterium intracellular larae (M. intracellulare), mycobacterium kansasii (M. kansasii), mycobacterium malmoense (M. malmoense), mycobacterium marinum (M. marinum), mycobacterium szulgai (M. szulgai) , Mycobacterium terrae (M. terrae), Mycobacterium scrofulaceum (M. scrofulaceum), Mycobacterium Ulcerans (M. ulcerans), Mycobacterium simiae (M. simiae) and Mycobacterium xenopi (M. xenopi) selected from the group consisting of, non-tuberculous mycobacterium infection or biomarker composition for diagnosis of infectious diseases.
  6. 제1항에 있어서,According to claim 1,
    상기 비결핵 항산균의 감염 질환은 폐 질환, 림프절염, 피부·연조직·골감염증 또는 파종성 질환인, 비결핵 항산균의 감염 또는 감염 질환의 진단용 바이오마커 조성물.The non-tuberculous mycobacterium infectious disease is a lung disease, lymphadenitis, skin, soft tissue, bone infection, or disseminated disease, a biomarker composition for diagnosis of non-tuberculous mycobacterium infection or infectious disease.
  7. 아미노산(amino acid), 아미노산 유도체, 알란토인(Allantoin), N,N-디메틸글라이신(N,N-Dimethylglycine), 2-하이드록시글루타릭산(2-hydroxyglutaric acid), 콜린(Choline), 하이포잔틴(Hypoxanthine), 락테이트(Lactate), 말산(Malic acid) 및 글리세롤 3-포스페이트(Glycerol 3-phosphate)로 이루어진 군에서 선택된 1종 이상의 대사체의 농도를 측정하는 정량 장치를 포함하는, 비결핵 항산균의 감염 또는 감염 질환의 진단용 키트.Amino acids, amino acid derivatives, allantoin, N,N-dimethylglycine, 2-hydroxyglutaric acid, choline, hypoxanthine ( Hypoxanthine), lactate (Lactate), malic acid (Malic acid) and glycerol 3-phosphate (Glycerol 3-phosphate) comprising a quantitative device for measuring the concentration of one or more metabolites selected from the group consisting of, non-tuberculous mycobacteria kit for the diagnosis of infections or infectious diseases.
  8. 제7항에 있어서,8. The method of claim 7,
    상기 아미노산 및 아미노산 유도체는 알기닌(Arginine), 프롤린(Proline), 트레오닌(Threonine), 류신(Leucine), 라이신(Lycine), 글루타메이트(Glutamate), 트립토판(Tryptophan), 아스파라긴(Asparagine), 메티오닌(Methionine), 세린(Serine), 호모세린(Homoserine) 및 타우린(Taurine)으로 이루어진 군에서 선택된 1종 이상을 포함하는, 비결핵 항산균의 감염 또는 감염 질환의 진단용 키트.The amino acids and amino acid derivatives are Arginine, Proline, Threonine, Leucine, Lysine, Glutamate, Tryptophan, Asparagine, Methionine. , Serine (Serine), homoserine (Homoserine) and taurine (Taurine), comprising at least one selected from the group consisting of, non-tuberculous mycobacterium infection or a diagnostic kit for infectious diseases.
  9. 제7항에 있어서,8. The method of claim 7,
    상기 아미노산은 L-형태(L-form)인, 비결핵 항산균의 감염 또는 감염 질환의 진단용 키트.The amino acid is L-form (L-form), non-tuberculous mycobacterium infection or infectious disease diagnostic kit.
  10. 제7항에 있어서, 8. The method of claim 7,
    상기 대사체는 목적하는 개체의 전혈(whole blood), 혈장(plasma) 또는 혈청(serum) 유래인 것인, 비결핵 항산균의 감염 또는 감염 질환의 진단용 키트.Wherein the metabolite is derived from whole blood, plasma or serum of a target individual, a kit for diagnosis of non-tuberculous mycobacteria infection or infectious disease.
  11. 제7항에 있어서, 8. The method of claim 7,
    상기 정량 장치는 핵자기 공명 분광 분석기 (NMR), 크로마토그래피 또는 질량분석기인, 비결핵 항산균의 감염 또는 감염 질환의 진단용 키트.The quantitative device is a nuclear magnetic resonance spectrometer (NMR), chromatography or mass spectrometer, a kit for diagnosing infection or infectious disease of non-tuberculous mycobacteria.
  12. 제7항에 있어서,8. The method of claim 7,
    상기 비결핵 항산균은 마이코박테리움 아비움(M. avium), 마이코박테리움 압세수스(M. abscessus), 마이코박테리움 플라베센스(M. flavescence), 마이코박테리움 아프리카눔(M. africanum), 마이코박테리움 보비스(M. bovis), 마이코박테리움 첼로네(M. chelonae), 마이코박테리움 셀라툼(M. celatum), 마이코박테리움 포르투이툼(M. fortuitum), 마이코박테리움 고르도네(M. gordonae), 마이코박테리움 가스트리(M. gastri), 마이코박테리움 헤모필룸(M. haemophilum), 마이코박테리움 인트라셀루라레(M. intracellulare), 마이코박테리움 칸사시이(M. kansasii), 마이코박테리움 말모엔스(M. malmoense), 마이코박테리움 마리눔(M. marinum), 마이코박테리움 스줄가이(M. szulgai), 마이코박테리움 테레(M. terrae), 마이코박테리움 스크로풀라세움(M. scrofulaceum), 마이코박테리움 울서란스(M. ulcerans), 마이코박테리움 시미애(M. simiae) 및 마이코박테리움 제노피(M. xenopi)로 구성된 군으로부터 선택되는, 비결핵 항산균의 감염 또는 감염 질환의 진단용 키트.The non-tuberculous mycobacteria are Mycobacterium avium (M. avium), Mycobacterium abscessus (M. abscessus), Mycobacterium flavescens (M. flavescence), Mycobacterium africanum (M. africanum), Mycobacterium bovis (M. bovis), Mycobacterium cellone (M. chelonae), Mycobacterium cellatum (M. celatum), Mycobacterium portuitum (M. fortuitum), Mycobacterium gordonae (M. gordonae), Mycobacterium gastri (M. gastri), Mycobacterium haemophilum (M. haemophilum), Mycobacterium intracellular larae (M. intracellulare), mycobacterium kansasii (M. kansasii), mycobacterium malmoense (M. malmoense), mycobacterium marinum (M. marinum), mycobacterium szulgai (M. szulgai) , Mycobacterium terrae (M. terrae), Mycobacterium scrofulaceum (M. scrofulaceum), Mycobacterium Ulcerans (M. ulcerans), Mycobacterium simiae (M. simiae) and Mycobacterium xenopi (M. xenopi) selected from the group consisting of, non-tuberculous mycobacterium infection or infectious disease diagnostic kit.
  13. 제7항에 있어서,8. The method of claim 7,
    상기 비결핵 항산균의 감염 질환은 폐 질환, 림프절염, 피부·연조직·골감염증 또는 파종성 질환인, 비결핵 항산균의 감염 또는 감염 질환의 진단용 키트.The non-tuberculous mycobacterium infectious disease is a lung disease, lymphadenitis, skin, soft tissue, bone infection, or disseminated disease, a kit for diagnosing an infection or an infectious disease of a non-tuberculous mycobacterium.
  14. 목적하는 개체로부터 분리된 생물학적 시료에서 아미노산(amino acid), 아미노산 유도체, 알란토인(Allantoin), N,N-디메틸글라이신(N,N-Dimethylglycine), 2-하이드록시글루타릭산(2-hydroxyglutaric acid), 콜린(Choline), 하이포잔틴(Hypoxanthine), 락테이트(Lactate), 말산(Malic acid) 및 글리세롤 3-포스페이트(Glycerol 3-phosphate)로 이루어진 군에서 선택된 1종 이상의 대사체의 발현 수준을 측정하는 단계를 포함하는 비결핵 항산균의 감염 또는 감염 질환을 진단하기 위한 정보 제공 방법.Amino acids, amino acid derivatives, allantoin, N,N-dimethylglycine, 2-hydroxyglutaric acid from a biological sample isolated from a subject of interest , Choline (Choline), hypoxanthine (Hypoxanthine), lactate (Lactate), malic acid (Malic acid) and glycerol 3-phosphate (Glycerol 3-phosphate) to measure the expression level of one or more metabolites selected from the group consisting of A method of providing information for diagnosing an infection or an infectious disease of a non-tuberculous mycobacterium comprising the steps of
  15. 제14항에 있어서,15. The method of claim 14,
    상기 생물학적 시료는 전혈(whole blood), 백혈구(leukocytes), 말초혈액 단핵 세포(peripheral blood mononuclear cells), 백혈구 연층(buffy coat), 혈장(plasma), 혈청(serum), 객담(sputum), 눈물(tears), 점액(mucus), 세비액(nasal washes), 비강 흡인물(nasal aspirate), 호흡(breath), 소변(urine), 정액(semen), 침(saliva), 복강 세척액(peritoneal washings), 복수(ascites), 낭종액(cystic fluid), 뇌척수막 액(meningeal fluid), 양수(amniotic fluid), 선액(glandular fluid), 췌장액(pancreatic fluid), 림프액(lymph fluid), 흉수(pleural fluid), 유두 흡인물(nipple aspirate), 기관지 흡인물(bronchial aspirate), 활액(synovial fluid), 관절 흡인물(joint aspirate), 기관 분비물(organ secretions), 세포(cell), 세포 추출물(cell extract) 및 뇌척수액(cerebrospinal fluid) 등으로 이루어진 군에서 선택된 1종 이상인, 비결핵 항산균의 감염 또는 감염 질환을 진단하기 위한 정보 제공 방법.The biological sample includes whole blood, leukocytes, peripheral blood mononuclear cells, buffy coat, plasma, serum, sputum, tears ( tears), mucus, nasal washes, nasal aspirate, breath, urine, semen, saliva, peritoneal washings, Ascites, cystic fluid, meningeal fluid, amniotic fluid, glandular fluid, pancreatic fluid, lymph fluid, pleural fluid, nipple aspirate, bronchial aspirate, synovial fluid, joint aspirate, organ secretions, cells, cell extract and cerebrospinal fluid ( cerebrospinal fluid) and the like, at least one selected from the group consisting of, non-tuberculous mycobacterium infection or an information providing method for diagnosing an infectious disease.
  16. 제14항에 있어서,15. The method of claim 14,
    상기 대사체의 발현 수준을 측정하기에 앞서, 상기 생물학적 시료를 여과, 증류, 추출, 분리, 농축, 방해 성분의 불활성화 또는 시약의 첨가의 전처리하는 단계를 더 포함하는, 비결핵 항산균의 감염 또는 감염 질환을 진단하기 위한 정보 제공 방법.Prior to measuring the expression level of the metabolite, the biological sample is pretreated with filtration, distillation, extraction, separation, concentration, inactivation of interfering components, or addition of a reagent. or a method of providing information for diagnosing an infectious disease.
  17. 제14항에 있어서,15. The method of claim 14,
    상기 아미노산 및 아미노산 유도체는 알기닌(Arginine), 프롤린(Proline), 트레오닌(Threonine), 류신(Leucine), 라이신(Lycine), 글루타메이트(Glutamate), 트립토판(Tryptophan), 아스파라긴(Asparagine), 메티오닌(Methionine), 세린(Serine), 호모세린(Homoserine) 및 타우린(Taurine)으로 이루어진 군에서 선택된 1종 이상을 포함하는, 비결핵 항산균의 감염 또는 감염 질환을 진단하기 위한 정보 제공 방법.The amino acids and amino acid derivatives are Arginine, Proline, Threonine, Leucine, Lysine, Glutamate, Tryptophan, Asparagine, Methionine. , Serine (Serine), homoserine (Homoserine) and taurine (Taurine), comprising at least one selected from the group consisting of, information providing method for diagnosing infection or infectious disease of non-tuberculous mycobacteria.
  18. 제14항에 있어서,15. The method of claim 14,
    상기 아미노산은 L-형태(L-form)인, 비결핵 항산균의 감염 또는 감염 질환을 진단하기 위한 정보 제공 방법.The amino acid is L-form (L-form), information providing method for diagnosing infection or infectious disease of non-tuberculous mycobacteria.
  19. 제14항에 있어서, 15. The method of claim 14,
    상기 대사체의 발현 수준을 측정하는 단계는 핵자기 공명 분광 분석기 (NMR), 크로마토그래피 또는 질량분석기인 정량 장치를 이용하여 수행되는, 비결핵 항산균의 감염 또는 감염 질환을 진단하기 위한 정보 제공 방법.The step of measuring the expression level of the metabolite is performed using a quantitative device that is a nuclear magnetic resonance spectrometer (NMR), chromatography, or mass spectrometer. Information providing method for diagnosing infection or infectious disease of non-tuberculous mycobacteria .
  20. 제17항에 있어서,18. The method of claim 17,
    상기 목적하는 개체의 생물학적 시료에 대하여 측정된 알기닌(Arginine) 및 알란토인(Allantoin) 중 적어도 하나의 발현 수준이 대조군에 비하여 증가된 경우, 비결핵 항산균에 의해 감염되었거나 감염 가능성이 높거나, 또는 감염 질환이 발병하였거나 발병 가능성이 높은 것으로 예측하는 단계를 더 포함하는, 비결핵 항산균의 감염 또는 감염 질환을 진단하기 위한 정보 제공 방법.When the expression level of at least one of arginine and allantoin measured with respect to the biological sample of the subject of interest is increased compared to the control group, it is infected by a non-tuberculous mycobacterium or has a high probability of infection, or infection A method of providing information for diagnosing an infection or an infectious disease of non-tuberculous mycobacteria, further comprising the step of predicting that the disease has occurred or is highly likely to develop.
  21. 제17항에 있어서,18. The method of claim 17,
    상기 목적하는 개체의 생물학적 시료에 대하여 측정된 프롤린(Proline), 트레오닌(Threonine), 류신(Leucine), 라이신(Lycine), 글루타메이트(Glutamate), 트립토판(Tryptophan), 아스파라긴(Asparagine), 메티오닌(Methionine), 세린(Serine), 호모세린(Homoserine), 타우린(Taurine), N,N-디메틸글라이신(N,N-Dimethylglycine), 2-하이드록시글루타릭산(2-hydroxyglutaric acid), 콜린(Choline), 하이포잔틴(Hypoxanthine), 락테이트(Lactate), 말산(Malic acid) 및 글리세롤 3-포스페이트(Glycerol 3-phosphate)으로 이루어진 군에서 선택된 1종 이상의 발현 수준이 대조군에 비하여 감소된 경우, 비결핵 항산균에 의해 감염되었거나 감염 가능성이 높거나, 또는 감염 질환이 발병하였거나 발병 가능성이 높은 것으로 예측하는 단계를 더 포함하는, 비결핵 항산균의 감염 또는 감염 질환을 진단하기 위한 정보 제공 방법.Proline, Threonine, Leucine, Lysine, Glutamate, Tryptophan, Asparagine, Methionine measured with respect to the biological sample of the subject of interest , Serine, Homoserine, Taurine, N,N-Dimethylglycine, 2-hydroxyglutaric acid, Choline, When the expression level of one or more selected from the group consisting of hypoxanthine, lactate, malic acid and glycerol 3-phosphate is reduced compared to the control group, non-tuberculous mycobacteria The method of providing information for diagnosing non-tuberculous mycobacteria infection or infectious disease, further comprising the step of predicting that the infection has been or is highly likely to be infected by, or an infectious disease has occurred or is highly likely to develop.
  22. 제14항에 있어서,15. The method of claim 14,
    상기 비결핵 항산균은 마이코박테리움 아비움(M. avium), 마이코박테리움 압세수스(M. abscessus), 마이코박테리움 플라베센스(M. flavescence), 마이코박테리움 아프리카눔(M. africanum), 마이코박테리움 보비스(M. bovis), 마이코박테리움 첼로네(M. chelonae), 마이코박테리움 셀라툼(M. celatum), 마이코박테리움 포르투이툼(M. fortuitum), 마이코박테리움 고르도네(M. gordonae), 마이코박테리움 가스트리(M. gastri), 마이코박테리움 헤모필룸(M. haemophilum), 마이코박테리움 인트라셀루라레(M. intracellulare), 마이코박테리움 칸사시이(M. kansasii), 마이코박테리움 말모엔스(M. malmoense), 마이코박테리움 마리눔(M. marinum), 마이코박테리움 스줄가이(M. szulgai), 마이코박테리움 테레(M. terrae), 마이코박테리움 스크로풀라세움(M. scrofulaceum), 마이코박테리움 울서란스(M. ulcerans), 마이코박테리움 시미애(M. simiae) 및 마이코박테리움 제노피(M. xenopi)로 구성된 군으로부터 선택되는, 비결핵 항산균의 감염 또는 감염 질환을 진단하기 위한 정보 제공 방법.The non-tuberculous mycobacteria are Mycobacterium avium (M. avium), Mycobacterium abscessus (M. abscessus), Mycobacterium flavescens (M. flavescence), Mycobacterium africanum (M. africanum), Mycobacterium bovis (M. bovis), Mycobacterium cellone (M. chelonae), Mycobacterium cellatum (M. celatum), Mycobacterium portuitum (M. fortuitum), Mycobacterium gordonae (M. gordonae), Mycobacterium gastri (M. gastri), Mycobacterium haemophilum (M. haemophilum), Mycobacterium intracellular larae (M. intracellulare), mycobacterium kansasii (M. kansasii), mycobacterium malmoense (M. malmoense), mycobacterium marinum (M. marinum), mycobacterium szulgai (M. szulgai) , Mycobacterium terrae (M. terrae), Mycobacterium scrofulaceum (M. scrofulaceum), Mycobacterium Ulcerans (M. ulcerans), Mycobacterium simiae (M. simiae) and Selected from the group consisting of Mycobacterium xenopi (M. xenopi), an information providing method for diagnosing an infection or an infectious disease of non-tuberculous mycobacteria.
  23. 제14항에 있어서,15. The method of claim 14,
    상기 비결핵 항산균의 감염 질환은 폐 질환, 림프절염, 피부·연조직·골감염증 또는 파종성 질환인, 비결핵 항산균의 감염 또는 감염 질환을 진단하기 위한 정보 제공 방법.The method of providing information for diagnosing an infection or an infectious disease of a non-tuberculous mycobacterium, wherein the non-tuberculous mycobacterium infectious disease is a lung disease, lymphadenitis, skin/soft tissue/oste infection or disseminated disease.
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