WO2021107550A1 - Biomarker for predicting whether or not positivity for bacteria is retained after nontuberculous mycobacterium infection - Google Patents

Biomarker for predicting whether or not positivity for bacteria is retained after nontuberculous mycobacterium infection Download PDF

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WO2021107550A1
WO2021107550A1 PCT/KR2020/016661 KR2020016661W WO2021107550A1 WO 2021107550 A1 WO2021107550 A1 WO 2021107550A1 KR 2020016661 W KR2020016661 W KR 2020016661W WO 2021107550 A1 WO2021107550 A1 WO 2021107550A1
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mycobacterium
infection
prognosis
predicting
tuberculous mycobacteria
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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 WO2021107550A1 publication Critical patent/WO2021107550A1/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
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N2800/00—Detection or diagnosis of diseases
    • G01N2800/56—Staging of a disease; Further complications associated with the disease

Definitions

  • the present invention relates to a prognosis after infection by non-tuberculous mycobacteria, in particular, a biomarker for predicting whether or not spontaneous bacterial negative transformation does not occur and persistent bacterial positivity occurs without appropriate treatment, and a kit or prediction method for the prediction is about
  • 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
  • An object of the present invention is to provide a biomarker composition for predicting whether or not spontaneous bacterial negative transformation does not occur without treatment, in particular, as a prognosis after infection with non-tuberculous mycobacteria, and whether bacteria positivity continues.
  • Another object of the present invention is to provide a kit for predicting the prognosis after infection with non-tuberculous mycobacteria, in particular, whether spontaneous bacterial negative transformation does not occur without treatment and bacterial positivity continues.
  • Another object of the present invention is to provide a method for predicting the prognosis after infection with non-tuberculous mycobacteria, in particular, whether or not spontaneous bacterial negative transformation does not occur without treatment and whether bacterial positivity continues.
  • biomarker composition for predicting prognosis after infection with non-tuberculous mycobacteria, including metabolites.
  • the term "prognosis” means determining whether or not treatment success, recurrence, metastasis, drug reactivity, resistance, etc., in the subject after infection with non-tuberculous mycobacteria, etc., but for the purpose of the present invention, the prognosis is With appropriate treatment after infection with Mycobacterium tuberculosis, for example, without antibiotic administration, it means whether or not spontaneous bacterial transformation does not occur and bacterial positivity persists.
  • the biomarker for predicting prognosis according to the present invention is a person with a relatively poor prognosis after infection with non-tuberculous mycobacterium, that is, a person who does not or is less likely to develop spontaneous bacterial negative transformation without appropriate treatment, a person with a relatively good prognosis, that is, an appropriate It is a substance that can distinguish and diagnose those who spontaneously develop or have a high probability of occurrence without treatment. After being infected with non-tuberculous mycobacteria, it is an appropriate treatment, for example, without antibiotic administration, spontaneous bacterial negative transformation does not occur and the bacteria continue to grow. metabolites showing an increase or decrease in a biological sample derived from a positive subject, preferably a blood metabolite.
  • the "treatment” refers to an approach for obtaining a beneficial or desirable clinical result, and for the purposes of the present invention, the beneficial or desired clinical result is, but not limited to, alleviation of symptoms, reduction of the scope of disease. , stabilizing (i.e. not exacerbating) the disease state, delaying or reducing the rate of disease progression, amelioration or temporary alleviation and amelioration (partial or total) of the disease state, whether detectable or undetectable. and may mean increasing survival compared to the expected survival rate in the absence of treatment.
  • Treatment refers to both therapeutic treatment and prophylactic or prophylactic measures. Such treatments include the treatment required for the disorder being prevented as well as the disorder that has already occurred.
  • “Palliating" a disease means that the extent and/or undesirable clinical signs of the disease state are reduced and/or the time course of progression is delayed or prolonged, compared to no treatment. means to lose
  • the treatment may be performed using an antibiotic, but is not limited thereto.
  • 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 term "bacterial negative” means a state in which non-tuberculous mycobacteria are not detected in a smear test or culture test of sputum, and as an example, 'negative' in which non-tuberculous mycobacteria are not detected in all three consecutive sputum smear tests. ', or may be a case of 'negative' in which non-tuberculous mycobacteria are not detected in one sputum culture test, but is not limited thereto.
  • the term "positive bacteria” means a state in which non-tuberculous mycobacteria are detected in a smear test or culture test of sputum. It may be determined as 'positive' or may be determined as 'positive' in which non-tuberculous mycobacteria are detected in one sputum culture test, but is not limited thereto.
  • the term "continuous" means a continuous predetermined period after infection with non-tuberculous mycobacterium or from the first diagnosis of infection with non-tuberculous mycobacterium, for example, continuous until treatment. period, or any period, specifically 1 month or more, 2 months or more, 3 months or more, 4 months or more, 5 months or more, 6 months or more, 7 months or more, 8 months or more, 9 months or more, 10 months or more , over 11 months, over 12 months, over 13 months, over 14 months, over 15 months, over 16 months, over 17 months, over 18 months, over 19 months, over 20 months, over 21 months, over 22 months, 23 It may mean more than one month, or more than 24 months, but is not limited thereto.
  • 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- Dimethyl glycine (N,N-Dimethylglycine), hypoxanthine (Hypoxanthine), lactate (Lactate), malic acid (Malic acid) and glycerol 3-phosphate (Glycerol 3-phosphate) may include at least one selected from the group consisting of have.
  • 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 acid and its derivatives are selected from the group consisting of valine, threonine, isoleucine, leucine, tryptophan, methionine and homoserine. It may include more than one species.
  • the amino acid may be in L-form, preferably valine, threonine, isoleucine, leucine, tryptophan, or methionine. is preferably in the L-form (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.
  • kits for predicting prognosis after infection with non-tuberculous mycobacteria 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- Dimethyl glycine (N,N-Dimethylglycine), hypoxanthine (Hypoxanthine), lactate (Lactate), malic acid (Malic acid) and glycerol 3-phosphate (Glycerol 3-phosphate) may include at least one selected from the group consisting of have.
  • 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 acid and its derivatives are selected from the group consisting of valine, threonine, isoleucine, leucine, tryptophan, methionine and homoserine. It may include more than one species.
  • the amino acid may be in L-form, preferably valine, threonine, isoleucine, leucine, tryptophan, or methionine. is preferably in the L-form (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.
  • it relates to a method for providing information for predicting the prognosis after infection with non-tuberculous mycobacterium, comprising measuring the expression level of a metabolite in a biological sample isolated from a target subject.
  • the "target subject” refers to an individual whose infection is uncertain by non-tuberculous mycobacterium, means an individual with a high probability of infection, or is infected or diagnosed as infected by non-tuberculous mycobacterium, but continues to be positive without antibiotic administration An individual whose occurrence is uncertain, which means an individual with a high probability of developing a bacterium.
  • 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- Dimethyl glycine (N,N-Dimethylglycine), hypoxanthine (Hypoxanthine), lactate (Lactate), malic acid (Malic acid) and glycerol 3-phosphate (Glycerol 3-phosphate) may include at least one selected from the group consisting of have.
  • 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 acid and its derivatives are selected from the group consisting of valine, threonine, isoleucine, leucine, tryptophan, methionine and homoserine. It may include more than one species.
  • the amino acid may be in L-form, preferably valine, threonine, isoleucine, leucine, tryptophan, or methionine. is preferably in the 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 prognosis after infection by non-tuberculous mycobacteria is preferably poor, preferably without antibiotic administration.
  • the method may further include predicting that the positivity persists or is highly likely to persist.
  • the expression level of one or more selected from the group consisting of the group is increased compared to the control group, the prognosis after infection by non-tuberculous mycobacteria is poor, preferably, without appropriate treatment, for example, antibiotic administration, bacterial negative charge does not occur, It may further include the step of predicting that the bacterium positivity continues or is highly likely to continue.
  • the method may further include predicting that the bacterial positivity continues or is highly likely to continue without appropriate treatment, for example, antibiotic administration.
  • control group is a normal control not infected with non-tuberculous mycobacteria, the median value of the patient population (or the average value of the patient) infected with non-tuberculous mycobacteria, or the prognosis after infection with non-tuberculous mycobacteria.
  • the prognosis after infection by non-tuberculous mycobacteria is poor as described above, and preferably, if it is predicted that the positivity continues or is highly likely to continue without administration of an appropriate treatment, for example, antibiotics, the It may further include the step of performing an appropriate treatment such as administration of a drug for the disease to the target subject, for example, 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.
  • 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 in a biological sample of a target individual, it is possible to simply, easily and accurately predict the prognosis after infection by non-tuberculous mycobacteria, in particular, the occurrence of fungal negativity. Accordingly, it is expected that appropriate treatment, such as an early decision on whether to treat antibiotics or not, will be possible.
  • the prognosis after infection by non-tuberculous mycobacteria is simple, easy and accurate, in particular, whether continuously showing positivity without appropriate treatment. predictable.
  • L-valine L-Valine
  • MAC Mycobacterium avium complex
  • Figure 2 is a comparison of the expression level of L-threonine (L-Threonine) in the serum samples of patients with Mycobacterium avium complex (MAC) infection before antibiotic treatment in an embodiment of the present invention and a patient who continuously develops bacteria. A graph is shown.
  • L-Threonine L-threonine
  • MAC Mycobacterium avium complex
  • FIG. 3 is a comparison of the expression level of L-isoleucine in serum samples of patients with Mycobacterium avium complex (MAC) infection before antibiotic treatment in one embodiment of the present invention and patients who continuously develop bacterium positivity. A graph is shown.
  • MAC Mycobacterium avium complex
  • L-Leucine L-leucine
  • MAC Mycobacterium avium complex
  • L-Methionine L-methionine
  • MAC Mycobacterium avium complex
  • L-tryptophan L-tryptophan
  • MAC Mycobacterium avium complex
  • N,N-dimethylglycine N,N-Dimethylglycine
  • MAC Mycobacterium avium complex
  • FIG. 8 is a graph comparing the expression level of homoserine in the serum sample of a patient with Mycobacterium avium complex (MAC) infection before antibiotic treatment in an embodiment of the present invention and a patient who continuously develops bacteria. it has been shown
  • FIG. 9 is a view showing the expression level of S-lactate in serum samples of patients with Mycobacterium avium complex (MAC) infection before antibiotic treatment in an embodiment of the present invention and patients who continuously developed bacteria. A comparison graph is shown.
  • MAC Mycobacterium avium complex
  • Figure 10 shows the expression level of glycerol 3-phosphate in the serum samples of patients with Mycobacterium avium complex (MAC) infection before antibiotic treatment in an embodiment of the present invention and patients who continuously developed bacteria. A graph comparing them is shown.
  • MAC Mycobacterium avium complex
  • FIG. 11 shows the expression level of glycerol L-malic acid (L-Malic acid) in serum samples of patients with Mycobacterium avium complex (MAC) infection before antibiotic treatment in one embodiment of the present invention and patients who continuously developed bacteria. A graph comparing them is shown.
  • L-Malic acid glycerol L-malic acid
  • MAC Mycobacterium avium complex
  • FIG. 12 is a graph comparing the expression level of hypoxanthine in serum samples of patients with Mycobacterium avium complex (MAC) infection before antibiotic treatment in one embodiment of the present invention and a patient who continuously develops bacteria. it has been shown
  • FIG. 13 is a graph comparing the expression level of Allantoin in the serum samples of patients with Mycobacterium avium complex (MAC) infection before antibiotic treatment in one embodiment of the present invention and patients who continuously developed bacteria. will be.
  • MAC Mycobacterium avium complex
  • the present invention relates to a method for providing information for predicting the prognosis after infection with non-tuberculous mycobacteria, comprising the step of 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
  • specific examples include amino acids, amino acid derivatives, allantoin, N,N- Dimethyl glycine (N,N-Dimethylglycine), hypoxanthine (Hypoxanthine), lactate (Lactate), malic acid (Malic acid) and glycerol 3-phosphate (Glycerol 3-phosphate) may include at least one selected from the group consisting of have.
  • the amino acid and its derivatives are one selected from the group consisting of valine, threonine, isoleucine, leucine, tryptophan, methionine and homoserine. may include more than one.
  • the lactate may be in the S-form, and the malic acid and the amino acid may be in the L-form.
  • Mycobacterium avium complex ( avium : 46, intracellulare : 50, total 96) collected from Samsung Hospital in Seoul for approximately 6 years from January 2012 to August 2016. 96 serum samples before antibiotic treatment and 30 serum samples from patients who were continuously positive but not worsened without antibiotic treatment were prepared.
  • sample quality control SQC
  • SQC sample quality control
  • Dx0 (Success & Fail) represents the expression level of each metabolite in the serum samples of 96 patients infected with Mycobacterium avium complex (MAC) before the start of antibiotic treatment, and Persistence is without antibiotic treatment.
  • the expression level of each metabolite is shown in the serum samples of 30 patients with persistent bacterial positivity.
  • L-valine, L-threonine, L-isoleucine, L-leucine, L-tryptophan, L-methionine (L-Methionine), homoserine (Homoserine), N,N-dimethylglycine (N,N-Dimethylglycine), S-lactate (S-Lactate), glycerol 3-phosphate (Glycerol 3-phosphate), L-Malic acid, hypoxanthine, and allantoin are biomarkers for determining whether to treat with antibiotics by predicting that bacteria will continue to be positive without antibiotic administration after infection with non-tuberculous mycobacteria. found that it can be used as
  • the present invention relates to a prognosis after infection by non-tuberculous mycobacteria, in particular, a biomarker for predicting whether or not spontaneous bacterial negative transformation does not occur and persistent bacterial positivity occurs without appropriate treatment, and a kit or prediction method for the prediction is about

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Abstract

The present invention relates to a biomarker, a prediction kit, or a prediction method, the biomarker for predicting whether or not positivity for bacteria is retained without spontaneous negativity for bacteria with no appropriate treatment, as a prognosis after nontuberculous mycobacterium infection by means of metabolites.

Description

비결핵 항산균에 의한 감염 후 균 양성의 지속 여부 예측용 바이오마커Biomarker for predicting whether or not bacterial positivity continues after infection by non-tuberculous mycobacteria
본 발명은 비결핵 항산균에 의한 감염 후 예후로, 특히는 적절한 치료 없이는 자발적 균 음전이 발생하지 않고 지속적 균 양성이 발생하는 지 여부를 예측하기 위한 바이오마커와, 상기 예측을 위한 키트 또는 예측 방법에 관한 것이다. The present invention relates to a prognosis after infection by non-tuberculous mycobacteria, in particular, a biomarker for predicting whether or not spontaneous bacterial negative transformation does not occur and persistent bacterial positivity occurs without appropriate treatment, and a kit or prediction method for the prediction is about
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 후 예후로, 특히는 치료 없이는 자발적 균 음전이 발생하지 않고 균 양성이 지속되는 지 여부를 예측하기 위한 바이오마커 조성물을 제공하고자 한다. An object of the present invention is to provide a biomarker composition for predicting whether or not spontaneous bacterial negative transformation does not occur without treatment, in particular, as a prognosis after infection with non-tuberculous mycobacteria, and whether bacteria positivity continues.
본 발명의 다른 목적은 비결핵 항산균의 감염 후 예후로, 특히는 치료 없이는 자발적 균 음전이 발생하지 않고 균 양성이 지속되는 지 여부를 예측하기 위한 키트를 제공하고자 한다.Another object of the present invention is to provide a kit for predicting the prognosis after infection with non-tuberculous mycobacteria, in particular, whether spontaneous bacterial negative transformation does not occur without treatment and bacterial positivity continues.
본 발명의 또 다른 목적은 비결핵 항산균의 감염 후 예후로, 특히는 치료 없이는 자발적 균 음전이 발생하지 않고 균 양성이 지속되는 지 여부를 예측하는 방법을 제공하고자 한다. Another object of the present invention is to provide a method for predicting the prognosis after infection with non-tuberculous mycobacteria, in particular, whether or not spontaneous bacterial negative transformation does not occur without treatment and whether bacterial positivity continues.
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 후 예후 예측용 바이오마커 조성물에 관한 것이다. According to one embodiment of the present invention, it relates to a biomarker composition for predicting prognosis after infection with non-tuberculous mycobacteria, including metabolites.
본 발명에서 "예후"란 비결핵 항산균의 감염 후 해당 개체에서 치료 성공 여부, 재발, 전이, 약물 반응성, 내성 등과 같은 여부를 판단하는 것을 의미하나, 본 발명의 목적 상 상기 상기 예후는 상기 비결핵 항산균의 감염 후 적절한 치료로, 예를 들면 항생제 투여 없이는 자발적 균 음전이 발생하지 않고 균 양성이 지속되는 지 여부를 의미한다.In the present invention, the term "prognosis" means determining whether or not treatment success, recurrence, metastasis, drug reactivity, resistance, etc., in the subject after infection with non-tuberculous mycobacteria, etc., but for the purpose of the present invention, the prognosis is With appropriate treatment after infection with Mycobacterium tuberculosis, for example, without antibiotic administration, it means whether or not spontaneous bacterial transformation does not occur and bacterial positivity persists.
본 발명에 따른 예후 예측용 바이오마커는 비결핵 항산균의 감염 후 예후가 상대적으로 나쁜 자, 즉 적절한 치료 없이는 자발적 균 음전이 발생하지 않거나 발생할 가능성이 낮은 자를, 예후가 상대적으로 좋은 자로, 즉 적절한 치료 없이도 자발적 균 음전이 발생하거나 발생할 가능성이 높은 자를 구분하여 진단할 수 있는 물질로, 비결핵 항산균에 의해 감염된 후 적절한 치료로, 예를 들면 항생제 투여 없이는 자발적 균 음전이 발생하지 않고 지속적으로 균 양성을 보이는 대상체 유래의 생물학적 시료에서 증가 또는 감소 양상을 보이는 대사체, 바람직하게는 혈액 대사체를 포함한다.The biomarker for predicting prognosis according to the present invention is a person with a relatively poor prognosis after infection with non-tuberculous mycobacterium, that is, a person who does not or is less likely to develop spontaneous bacterial negative transformation without appropriate treatment, a person with a relatively good prognosis, that is, an appropriate It is a substance that can distinguish and diagnose those who spontaneously develop or have a high probability of occurrence without treatment. After being infected with non-tuberculous mycobacteria, it is an appropriate treatment, for example, without antibiotic administration, spontaneous bacterial negative transformation does not occur and the bacteria continue to grow. metabolites showing an increase or decrease in a biological sample derived from a positive subject, preferably a blood metabolite.
본 발명에서 상기 "치료"는 이롭거나 바람직한 임상적 결과를 수득하기 위한 접근을 의미하는 것으로, 본 발명의 목적을 위해서, 이롭거나 바람직한 임상적 결과는 비제한적으로, 증상의 완화, 질병 범위의 감소, 질병 상태의 안정화 (즉, 악화되지 않음), 질병 진행의 지연 또는 속도의 감소, 질병 상태의 개선 또는 일시적 완화 및 경감 (부분적이거나 전체적으로), 검출 가능 하거나 또는 검출되지 않거나의 여부를 포함할 수 있고, 치료를 받지 않았을 때 예상되는 생존율과 비교하여 생존율을 늘이는 것을 의미할 수도 있다. 치료는 치료학적 치료 및 예방적 또는 예방 조치 방법 모두를 가리킨다. 상기 치료들은 예방되는 장애뿐만 아니라 이미 발생한 장애에 있어서 요구되는 치료를 포함한다. 질병을 "완화(Palliating)"하는 것은 치료를 하지 않은 경우와 비교하여, 질병 상태의 범위 및/또는 바람직하지 않은 임상적 징후가 감소되거나 및/또는 진행의 시간적 추이(time course)가 늦춰지거나 길어지는 것을 의미한다. 본 발명의 목적상 상기 치료는 항생제를 사용하여 수행될 수 있으나, 이에 제한되는 것은 아니다. In the present invention, the "treatment" refers to an approach for obtaining a beneficial or desirable clinical result, and for the purposes of the present invention, the beneficial or desired clinical result is, but not limited to, alleviation of symptoms, reduction of the scope of disease. , stabilizing (i.e. not exacerbating) the disease state, delaying or reducing the rate of disease progression, amelioration or temporary alleviation and amelioration (partial or total) of the disease state, whether detectable or undetectable. and may mean increasing survival compared to the expected survival rate in the absence of treatment. Treatment refers to both therapeutic treatment and prophylactic or prophylactic measures. Such treatments include the treatment required for the disorder being prevented as well as the disorder that has already occurred. "Palliating" a disease means that the extent and/or undesirable clinical signs of the disease state are reduced and/or the time course of progression is delayed or prolonged, compared to no treatment. means to lose For the purposes of the present invention, the treatment may be performed using an antibiotic, but is not limited thereto.
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 발명에서 상기 "균 음전"이란 객담의 도말 검사 또는 배양 검사 상 비결핵 항산균이 검출되지 않은 상태를 의미하며, 일 예시로 객담 도말 검사 연속 3회 모두 비결핵 항산균이 검출되지 않는 '음성'으로 판정되거나, 객담 배양 검사 1회에서 비결핵 항산균이 검출되지 않는 '음성'으로 판정된 경우일 수 있으나, 이에 제한되는 것은 아니다. In the present invention, the term "bacterial negative" means a state in which non-tuberculous mycobacteria are not detected in a smear test or culture test of sputum, and as an example, 'negative' in which non-tuberculous mycobacteria are not detected in all three consecutive sputum smear tests. ', or may be a case of 'negative' in which non-tuberculous mycobacteria are not detected in one sputum culture test, but is not limited thereto.
본 발명에서 상기 "균 양성"이란 객담의 도말 검사 또는 배양 검사 상 비결핵 항산균이 검출되는 상태를 의미하며, 일 예시로 객담 도말 검사 3회 중 어느 한 번이라도 비결핵 항산균이 검출되는 '양성'으로 판정되거나, 객담 배양 검사 1회에서 비결핵 항산균이 검출되는 '양성'으로 판정된 경우일 수 있으나, 이에 제한되는 것은 아니다. In the present invention, the term "positive bacteria" means a state in which non-tuberculous mycobacteria are detected in a smear test or culture test of sputum. It may be determined as 'positive' or may be determined as 'positive' in which non-tuberculous mycobacteria are detected in one sputum culture test, but is not limited thereto.
본 발명에서 상기 "지속"의 의미는 비결핵 항산균의 감염 후 또는 비결핵 항산균에 의해 감염된 것으로 처음 진단받은 날로부터 연속된 소정의 기간을 의미하는 것으로, 예를 들면, 치료 전까지의 연속된 기간이거나, 임의의 기간으로, 구체적으로는 1 개월 이상, 2 개월 이상, 3 개월 이상, 4 개월 이상, 5 개월 이상, 6 개월 이상, 7 개월 이상, 8 개월 이상, 9 개월 이상, 10 개월 이상, 11 개월 이상, 12 개월 이상, 13 개월 이상, 14 개월 이상, 15 개월 이상, 16 개월 이상, 17 개월 이상, 18 개월 이상, 19 개월 이상, 20 개월 이상, 21 개월 이상, 22 개월 이상, 23 개월 이상, 또는 24 개월 이상을 의미할 수 있으나, 이에 제한되는 것은 아니다. In the present invention, the term "continuous" means a continuous predetermined period after infection with non-tuberculous mycobacterium or from the first diagnosis of infection with non-tuberculous mycobacterium, for example, continuous until treatment. period, or any period, specifically 1 month or more, 2 months or more, 3 months or more, 4 months or more, 5 months or more, 6 months or more, 7 months or more, 8 months or more, 9 months or more, 10 months or more , over 11 months, over 12 months, over 13 months, over 14 months, over 15 months, over 16 months, over 17 months, over 18 months, over 19 months, over 20 months, over 21 months, over 22 months, 23 It may mean more than one month, or more than 24 months, but is not limited thereto.
본 발명에서 상기 "대사체(metabolite)"는 대사물질 또는 대사산물이라고도 불리우며, 물질 대사의 중간생성물 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), 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- Dimethyl glycine (N,N-Dimethylglycine), hypoxanthine (Hypoxanthine), lactate (Lactate), malic acid (Malic acid) and glycerol 3-phosphate (Glycerol 3-phosphate) may include at least one selected from the group consisting of have.
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 발린(Valine), 트레오닌(Threonine), 이소류신(Isoleucine), 류신(Leucine), 트립토판(Tryptophan), 메티오닌(Methionine) 및 호모세린(Homoserine)으로 이루어진 군에서 선택된 1종 이상을 포함할 수 있다. In the present invention, the amino acid and its derivatives are selected from the group consisting of valine, threonine, isoleucine, leucine, tryptophan, methionine and homoserine. It may include more than one species.
본 발명에서 상기 아미노산은 L-형태(L-form)일 수 있고, 바람직하게는 발린(Valine), 트레오닌(Threonine), 이소류신(Isoleucine), 류신(Leucine), 트립토판(Tryptophan) 또는 메티오닌(Methionine)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 valine, threonine, isoleucine, leucine, tryptophan, or methionine. is preferably in the L-form (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 발명의 다른 구현 예에 따르면, 대사체의 농도를 측정하는 정량 장치를 포함하는, 비결핵 항산균의 감염 후 예후 예측용 키트에 관한 것이다. According to another embodiment of the present invention, it relates to a kit for predicting prognosis after infection with non-tuberculous mycobacteria, 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), 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- Dimethyl glycine (N,N-Dimethylglycine), hypoxanthine (Hypoxanthine), lactate (Lactate), malic acid (Malic acid) and glycerol 3-phosphate (Glycerol 3-phosphate) may include at least one selected from the group consisting of have.
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 발린(Valine), 트레오닌(Threonine), 이소류신(Isoleucine), 류신(Leucine), 트립토판(Tryptophan), 메티오닌(Methionine) 및 호모세린(Homoserine)으로 이루어진 군에서 선택된 1종 이상을 포함할 수 있다. In the present invention, the amino acid and its derivatives are selected from the group consisting of valine, threonine, isoleucine, leucine, tryptophan, methionine and homoserine. It may include more than one species.
본 발명에서 상기 아미노산은 L-형태(L-form)일 수 있고, 바람직하게는 발린(Valine), 트레오닌(Threonine), 이소류신(Isoleucine), 류신(Leucine), 트립토판(Tryptophan) 또는 메티오닌(Methionine)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 valine, threonine, isoleucine, leucine, tryptophan, or methionine. is preferably in the L-form (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 항산균 및 예후에 관한 정의는 상기 본 발명의 바이오마커 조성물에 기재된 바와 중복되어 명세서의 과도한 혼잡을 피하기 위해 이하 그 기재를 생략한다. Definitions regarding non-tuberculous mycobacteria and prognosis in the kit of the present invention overlap with those described in the biomarker composition of the present invention, and description thereof will be omitted below to avoid excessive confusion of the specification.
본 발명의 또 다른 구현 예에 따르면, 목적하는 개체로부터 분리된 생물학적 시료에서 대사체의 발현 수준을 측정하는 단계를 포함하는 비결핵 항산균의 감염 후 예후를 예측하기 위한 정보 제공 방법에 관한 것이다. According to another embodiment of the present invention, it relates to a method for providing information for predicting the prognosis after infection with non-tuberculous mycobacterium, 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 의미하거나, 비결핵 항산균에 의해 감염되었거나 감염된 것으로 진단받았으나 항생제 투여 없이는 지속적 균 양성 발생 여부가 불확실한 개체로, 균 양성 발생 가능성이 높은 개체를 의미한다.In the present invention, the "target subject" refers to an individual whose infection is uncertain by non-tuberculous mycobacterium, means an individual with a high probability of infection, or is infected or diagnosed as infected by non-tuberculous mycobacterium, but continues to be positive without antibiotic administration An individual whose occurrence is uncertain, which means an individual with a high probability of developing a bacterium.
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), 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- Dimethyl glycine (N,N-Dimethylglycine), hypoxanthine (Hypoxanthine), lactate (Lactate), malic acid (Malic acid) and glycerol 3-phosphate (Glycerol 3-phosphate) may include at least one selected from the group consisting of have.
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 발린(Valine), 트레오닌(Threonine), 이소류신(Isoleucine), 류신(Leucine), 트립토판(Tryptophan), 메티오닌(Methionine) 및 호모세린(Homoserine)으로 이루어진 군에서 선택된 1종 이상을 포함할 수 있다. In the present invention, the amino acid and its derivatives are selected from the group consisting of valine, threonine, isoleucine, leucine, tryptophan, methionine and homoserine. It may include more than one species.
본 발명에서 상기 아미노산은 L-형태(L-form)일 수 있고, 바람직하게는 발린(Valine), 트레오닌(Threonine), 이소류신(Isoleucine), 류신(Leucine), 트립토판(Tryptophan) 또는 메티오닌(Methionine)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 valine, threonine, isoleucine, leucine, tryptophan, or methionine. is preferably in the 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 의한 감염 후 예후가 나쁠 것, 바람직하게는 항생제 투여 없이는 균 양성이 지속되거나 지속될 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, the prognosis after infection by non-tuberculous mycobacteria is preferably poor, preferably without antibiotic administration. The method may further include predicting that the positivity persists or is highly likely to persist.
본 발명의 일 예시로, 본 발명에서 상기 목적하는 개체의 생물학적 시료에 대하여 측정된 발린(Valine), 트레오닌(Threonine), 이소류신(Isoleucine), 류신(Leucine), 트립토판(Tryptophan), 메티오닌(Methionine), 호모세린(Homoserine), N,N-디메틸글라이신(N,N-Dimethylglycine), 하이포잔틴(Hypoxanthine), 락테이트(Lactate), 말산(Malic acid) 및 글리세롤 3-포스페이트(Glycerol 3-phosphate)로 이루어진 군에서 선택된 1종 이상의 발현 수준이 대조군에 비하여 증가된 경우, 비결핵 항산균에 의한 감염 후 예후가 나쁠 것, 바람직하게는 적절한 치료, 예를 들면, 항생제 투여 없이는 균 음전이 발생하지 않고, 균 양성이 지속되거나 지속될 가능성이 높은 것으로 예측하는 단계를 더 포함할 수 있다.As an example of the present invention, in the present invention, valine, threonine, isoleucine, leucine, tryptophan, methionine measured with respect to the biological sample of the subject of interest in the present invention , homoserine, N,N-dimethylglycine (N,N-Dimethylglycine), hypoxanthine, lactate (Lactate), malic acid (Malic acid) and glycerol 3-phosphate (Glycerol 3-phosphate) When the expression level of one or more selected from the group consisting of the group is increased compared to the control group, the prognosis after infection by non-tuberculous mycobacteria is poor, preferably, without appropriate treatment, for example, antibiotic administration, bacterial negative charge does not occur, It may further include the step of predicting that the bacterium positivity continues or is highly likely to continue.
본 발명의 다른 예시로, 본 발명에서 상기 목적하는 개체의 생물학적 시료에 대하여 측정된 알란토인(Allantoin)의 발현 수준이 대조군에 비하여 감소된 경우, 비결핵 항산균에 의한 감염 후 예후가 나쁠 것, 바람직하게는 적절한 치료, 예를 들면, 항생제 투여 없이는 균 양성이 지속되거나 지속될 가능성이 높은 것으로 예측하는 단계를 더 포함할 수 있다.As another example of the present invention, when the expression level of Allantoin measured with respect to the biological sample of the subject of the present invention is reduced compared to the control, the prognosis after infection by non-tuberculous mycobacteria is poor, preferably For example, the method may further include predicting that the bacterial positivity continues or is highly likely to continue without appropriate treatment, for example, antibiotic administration.
본 발명에서 상기 "대조군"이란 비결핵 항산균에 의해 감염되지 않은 정상 대조군이거나, 비결핵 항산균에 의해 감염된 환자 모집단의 중앙값(또는 해당 환자의 평균값)이거나, 비결핵 항산균에 의한 감염 후 예후가 좋은 환자, 바람직하게는 균 음전이 발생하거나 발생할 가능성이 높은 환자, 보다 바람직하게는 적절한 치료 없이도 자발적으로 균 음전이 발생하거나 발생할 가능성이 높은 환자 모집단의 중앙값(또는 해당 환자의 평균값)일 수 있다.In the present invention, the "control group" is a normal control not infected with non-tuberculous mycobacteria, the median value of the patient population (or the average value of the patient) infected with non-tuberculous mycobacteria, or the prognosis after infection with non-tuberculous mycobacteria. may be the median (or mean value of those patients) of a population of good patients, preferably those who develop or are likely to develop, more preferably, those who develop or are likely to develop spontaneously, even without appropriate treatment. .
더 나아가, 본 발명에서 상기와 같이 비결핵 항산균에 의한 감염 후 예후가 나쁠 것, 바람직하게는 적절한 치료, 예를 들면, 항생제 투여 없이는 균 양성이 지속되거나 지속될 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, the prognosis after infection by non-tuberculous mycobacteria is poor as described above, and preferably, if it is predicted that the positivity continues or is highly likely to continue without administration of an appropriate treatment, for example, antibiotics, the It may further include the step of performing an appropriate treatment such as administration of a drug for the disease to the target subject, for example, 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 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.
이처럼 본 발명에서는 목적하는 개체의 생물학적 시료에 대하여 혈액 대사체의 발현 수준을 측정함으로써 비결핵 항산균에 의한 감염 후 예후로, 특히는 균 음전의 발생 여부를 간단하고 용이하면서도 정확하게 예측할 수 있고, 이에 따라 항생제 치료 여부를 조기에 결정하는 등 적절한 치료를 할 수 있을 것으로 기대된다.As such, in the present invention, by measuring the expression level of a blood metabolite in a biological sample of a target individual, it is possible to simply, easily and accurately predict the prognosis after infection by non-tuberculous mycobacteria, in particular, the occurrence of fungal negativity. Accordingly, it is expected that appropriate treatment, such as an early decision on whether to treat antibiotics or not, will be possible.
본 발명에서는 목적하는 개체의 생물학적 시료에 대하여 혈액 대사체의 발현 수준을 측정함으로써 비결핵 항산균에 의한 감염 후 예후로, 특히는 적절한 치료 없이는 지속적으로 균 양성을 보이는 지 여부를 간단하고 용이하면서도 정확하게 예측할 수 있다.In the present invention, by measuring the expression level of blood metabolites in a biological sample of a target individual, the prognosis after infection by non-tuberculous mycobacteria is simple, easy and accurate, in particular, whether continuously showing positivity without appropriate treatment. predictable.
도 1은 본 발명의 일 실시예에서 항생제 치료 전의 마이코박테리움 아비움 복합체(MAC) 감염 환자와 지속적으로 균 양성이 발생한 환자의 혈청 시료에서 L-발린(L-Valine)의 발현 수준을 비교한 그래프를 나타낸 것이다.1 is a comparison of the expression level of L-valine (L-Valine) in the serum sample of a patient with Mycobacterium avium complex (MAC) infection before antibiotic treatment in an embodiment of the present invention and a patient who continuously develops bacteria. A graph is shown.
도 2는 본 발명의 일 실시예에서 항생제 치료 전의 마이코박테리움 아비움 복합체(MAC) 감염 환자와 지속적으로 균 양성이 발생한 환자의 혈청 시료에서 L-트레오닌(L-Threonine)의 발현 수준을 비교한 그래프를 나타낸 것이다.Figure 2 is a comparison of the expression level of L-threonine (L-Threonine) in the serum samples of patients with Mycobacterium avium complex (MAC) infection before antibiotic treatment in an embodiment of the present invention and a patient who continuously develops bacteria. A graph is shown.
도 3은 본 발명의 일 실시예에서 항생제 치료 전의 마이코박테리움 아비움 복합체(MAC) 감염 환자와 지속적으로 균 양성이 발생한 환자의 혈청 시료에서 L-이소류신(L-Isoleucine)의 발현 수준을 비교한 그래프를 나타낸 것이다.3 is a comparison of the expression level of L-isoleucine in serum samples of patients with Mycobacterium avium complex (MAC) infection before antibiotic treatment in one embodiment of the present invention and patients who continuously develop bacterium positivity. A graph is shown.
도 4는 본 발명의 일 실시예에서 항생제 치료 전의 마이코박테리움 아비움 복합체(MAC) 감염 환자와 지속적으로 균 양성이 발생한 환자의 혈청 시료에서 L-류신(L-Leucine)의 발현 수준을 비교한 그래프를 나타낸 것이다.4 is a comparison of the expression level of L-leucine (L-Leucine) in the serum samples of patients with Mycobacterium avium complex (MAC) infection before antibiotic treatment in an embodiment of the present invention and patients who continuously developed bacteria. A graph is shown.
도 5는 본 발명의 일 실시예에서 항생제 치료 전의 마이코박테리움 아비움 복합체(MAC) 감염 환자와 지속적으로 균 양성이 발생한 환자의 혈청 시료에서 L-메티오닌(L-Methionine)의 발현 수준을 비교한 그래프를 나타낸 것이다.5 is a comparison of the expression level of L-methionine (L-Methionine) in the serum samples of patients with Mycobacterium avium complex (MAC) infection before antibiotic treatment in an embodiment of the present invention and a patient who continuously develops bacteria. A graph is shown.
도 6은 본 발명의 일 실시예에서 항생제 치료 전의 마이코박테리움 아비움 복합체(MAC) 감염 환자와 지속적으로 균 양성이 발생한 환자의 혈청 시료에서 L-트립토판(L-Tryptophan)의 발현 수준을 비교한 그래프를 나타낸 것이다.6 is a comparison of the expression level of L-tryptophan (L-Tryptophan) in the serum samples of patients with Mycobacterium avium complex (MAC) infection before antibiotic treatment in an embodiment of the present invention and a patient who continuously develops bacteria. A graph is shown.
도 7은 본 발명의 일 실시예에서 항생제 치료 전의 마이코박테리움 아비움 복합체(MAC) 감염 환자와 지속적으로 균 양성이 발생한 환자의 혈청 시료에서 N,N-디메틸글라이신(N,N-Dimethylglycine)의 발현 수준을 비교한 그래프를 나타낸 것이다.7 is N,N-dimethylglycine (N,N-Dimethylglycine) in the serum sample of a patient with Mycobacterium avium complex (MAC) infection before antibiotic treatment in an embodiment of the present invention and a patient who continuously develops bacteria. It shows a graph comparing the expression level of
도 8은 본 발명의 일 실시예에서 항생제 치료 전의 마이코박테리움 아비움 복합체(MAC) 감염 환자와 지속적으로 균 양성이 발생한 환자의 혈청 시료에서 호모세린(Homoserine)의 발현 수준을 비교한 그래프를 나타낸 것이다.8 is a graph comparing the expression level of homoserine in the serum sample of a patient with Mycobacterium avium complex (MAC) infection before antibiotic treatment in an embodiment of the present invention and a patient who continuously develops bacteria. it has been shown
도 9는 본 발명의 일 실시예에서 항생제 치료 전의 마이코박테리움 아비움 복합체(MAC) 감염 환자와 지속적으로 균 양성이 발생한 환자의 혈청 시료에서 S-락테이트(S-Lactate)의 발현 수준을 비교한 그래프를 나타낸 것이다.9 is a view showing the expression level of S-lactate in serum samples of patients with Mycobacterium avium complex (MAC) infection before antibiotic treatment in an embodiment of the present invention and patients who continuously developed bacteria. A comparison graph is shown.
도 10은 본 발명의 일 실시예에서 항생제 치료 전의 마이코박테리움 아비움 복합체(MAC) 감염 환자와 지속적으로 균 양성이 발생한 환자의 혈청 시료에서 글리세롤 3-포스페이트(Glycerol 3-phosphate)의 발현 수준을 비교한 그래프를 나타낸 것이다.Figure 10 shows the expression level of glycerol 3-phosphate in the serum samples of patients with Mycobacterium avium complex (MAC) infection before antibiotic treatment in an embodiment of the present invention and patients who continuously developed bacteria. A graph comparing them is shown.
도 11은 본 발명의 일 실시예에서 항생제 치료 전의 마이코박테리움 아비움 복합체(MAC) 감염 환자와 지속적으로 균 양성이 발생한 환자의 혈청 시료에서 글리세롤 L-말산(L-Malic acid)의 발현 수준을 비교한 그래프를 나타낸 것이다.11 shows the expression level of glycerol L-malic acid (L-Malic acid) in serum samples of patients with Mycobacterium avium complex (MAC) infection before antibiotic treatment in one embodiment of the present invention and patients who continuously developed bacteria. A graph comparing them is shown.
도 12는 본 발명의 일 실시예에서 항생제 치료 전의 마이코박테리움 아비움 복합체(MAC) 감염 환자와 지속적으로 균 양성이 발생한 환자의 혈청 시료에서 하이포잔틴(Hypoxanthine)의 발현 수준을 비교한 그래프를 나타낸 것이다.12 is a graph comparing the expression level of hypoxanthine in serum samples of patients with Mycobacterium avium complex (MAC) infection before antibiotic treatment in one embodiment of the present invention and a patient who continuously develops bacteria. it has been shown
도 13은 본 발명의 일 실시예에서 항생제 치료 전의 마이코박테리움 아비움 복합체(MAC) 감염 환자와 지속적으로 균 양성이 발생한 환자의 혈청 시료에서 알란토인(Allantoin)의 발현 수준을 비교한 그래프를 나타낸 것이다.13 is a graph comparing the expression level of Allantoin in the serum samples of patients with Mycobacterium avium complex (MAC) infection before antibiotic treatment in one embodiment of the present invention and patients who continuously developed bacteria. will be.
본 발명의 일 구현 예에 따르면, 목적하는 개체로부터 분리된 생물학적 시료에서 대사체의 발현 수준을 측정하는 단계를 포함하는 비결핵 항산균의 감염 후 예후를 예측하기 위한 정보 제공 방법에 관한 것이다. According to one embodiment of the present invention, it relates to a method for providing information for predicting the prognosis after infection with non-tuberculous mycobacteria, comprising the step of 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), 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 발린(Valine), 트레오닌(Threonine), 이소류신(Isoleucine), 류신(Leucine), 트립토판(Tryptophan), 메티오닌(Methionine) 및 호모세린(Homoserine)으로 이루어진 군에서 선택된 1종 이상을 포함할 수 있다. 또한, 본 발명에서 상기 락테이트(Lactate)는 S-형태(S-form)이고, 상기 말산(Malic acid) 및 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- Dimethyl glycine (N,N-Dimethylglycine), hypoxanthine (Hypoxanthine), lactate (Lactate), malic acid (Malic acid) and glycerol 3-phosphate (Glycerol 3-phosphate) may include at least one selected from the group consisting of have. Here, the amino acid and its derivatives are one selected from the group consisting of valine, threonine, isoleucine, leucine, tryptophan, methionine and homoserine. may include more than one. Also, in the present invention, the lactate may be in the S-form, and the malic acid and the amino acid may be in the 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 자발적으로 균 음전된 환자의 시료 수집[Experimental Example 1] Sample collection of MAC-infected patients and spontaneously negatively charged patients
2012년 1월부터 2016년 8월까지의 대략 6년간 서울 삼성병원에서 수집한 마이코박테리움 아비움 복합체(Mycobacterium avium complex) (avium : 46명, intracellulare : 50명, 총 96명) 감염 환자의 항생제 치료 전 혈청 샘플 96개와 항생제 치료 없이는 지속적으로 균 양성 상태이지만 악화되지 않은 환자의 혈청 샘플 30개를 준비하였다. Mycobacterium avium complex ( avium : 46, intracellulare : 50, total 96) collected from Samsung Hospital in Seoul for approximately 6 years from January 2012 to August 2016. 96 serum samples before antibiotic treatment and 30 serum samples from patients who were continuously positive but not worsened without antibiotic treatment were prepared.
[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 및 3과 같이 수행하였다. 단, 하기 표 2 및 3에서 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 산출하였다.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 and 3 below. However, in Tables 2 and 3, 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
포름산 방법 (+), 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 (-), 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)-LactateLactate (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
(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
(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 감염 후 자발적 균 음전 발생자의 혈청 시료 내 대사체 분석 결과[Experimental Example 4] Metabolite analysis results in serum samples of spontaneous bacterial negative electrons after infection with non-tuberculous mycobacteria
항생제 치료 전의 마이코박테리움 아비움 복합체(MAC) 감염 환자와 항생제 투여 없이 지속적으로 균 양성을 보인 환자의 대사체 농도를 비교하기 위해 다음의 통계 검정 2가지 방법으로 Metaboanalyst(data 통계사이트)와 SPSS 통계 프로그램을 이용하여 데이터를 산출하였고, 그 결과를 이용하여 항생제 투여 없이는 지속적 균 양성의 발생 여부를 예측할 수 있고 그에 따라 항생제 치료의 필요 여부를 결정할 수 있는 대사체 총 13 종(p-value>0.05)을 각각의 p-value와 발현 수준의 배수 변화(Fold change) 값을 토대로 선정하여 그 결과를 하기 표 4 및 5와, 도 1 내지 13에 나타내었다. 단, 도 1 내지 13에서 Dx0(Success & Fail)는 항생제 치료 시작 전의 마이코박테리움 아비움 복합체(MAC) 감염 환자 96명의 혈청 샘플에서 각 대사체의 발현 수준을 나타낸 것이고, Persistence는 항생제 치료 없이는 지속적으로 균 양성이 발생하는 환자 30명의 혈청 샘플에서 각 대사체의 발현 수준을 나타낸 것이다. 또한, 유의성 unpaired t-test에서 *P<0.05; **P<0.01; ***P<0.001를 의미한다. In order to compare the metabolite concentration of patients with Mycobacterium avium complex (MAC) infection before antibiotic treatment and patients who were continuously positive without antibiotic treatment, Metaboanalyst (data statistics site) and SPSS were used with the following two statistical tests. The data were calculated using a statistical program, and using the results, it is possible to predict the occurrence of persistent bacterial positivity without antibiotic administration, and accordingly, a total of 13 metabolites (p-value >0.05) that can determine the need for antibiotic treatment. ) was selected based on each p-value and the fold change value of the expression level, and the results are shown in Tables 4 and 5 and FIGS. 1 to 13 below. However, in FIGS. 1 to 13, Dx0 (Success & Fail) represents the expression level of each metabolite in the serum samples of 96 patients infected with Mycobacterium avium complex (MAC) before the start of antibiotic treatment, and Persistence is without antibiotic treatment. The expression level of each metabolite is shown in the serum samples of 30 patients with persistent bacterial positivity. In addition, *P<0.05 in the significance unpaired t-test; **P<0.01; *** means P<0.001.
자발적 균 음전 성공 환자 시료에서 증가된 대사체Increased metabolites in patient samples with spontaneous bacterial negative success
대사체 종류(Compounds)Metabolites (Compounds) 유의성 (p-value)Significance (p-value) Fold ChangeFold Change
발린(L-Valine)L-Valine 0.01150.0115 1.221.22
트레오닌(L-Threonine)Threonine (L-Threonine) 0.04380.0438 1.321.32
이소류신(L-isoLeucine)Isoleucine (L-isoLeucine) 0.04110.0411 1.521.52
류신(L-Leucine)Leucine (L-Leucine) 0.02730.0273 1.451.45
메티오닌(L-Methionine)Methionine (L-Methionine) 0.00440.0044 1.631.63
트립토판(L-Tryptophan)Tryptophan (L-Tryptophan) 0.00420.0042 1.321.32
N,N-다이메틸글라이신
(N,N-Diemthylglycine)N,N-dimethylglycine
(N,N-Diemthylglycine) 		0.02510.0251 1.311.31
호모세린(Homoserine)Homoserine 0.04160.0416 1.351.35
(S)-락테이트((S)-Lactate)(S)-Lactate ((S)-Lactate) 0.02960.0296 1.331.33
글리세롤 3-포스페이트
Glycerol 3-phosphateGlycerol 3-phosphate
Glycerol 3-phosphate 		0.04360.0436 2.232.23
하이포잔틴(Hypoxanthine)Hypoxanthine 0.05740.0574 1.311.31
말릭 엑시드(L-Malic acid)Malic acid (L-Malic acid) 0.08870.0887 2.072.07
자발적 균 음전 성공 환자 시료에서 감소되어 있는 대사체Reduced metabolites in patient samples with spontaneous bacterial negative success
대사체 종류(Compounds)Metabolites (Compounds) 유의성 (p-value)Significance (p-value) Fold ChangeFold Change
알란토인(Allantoin)Allantoin 0.00270.0027 0.860.86
상기 표 4 및 5와, 도 1 내지 13에서 보는 바와 같이, 혈액 대사체 중 L-발린(L-Valine), L-트레오닌(L-Threonine), L-이소류신(L-Isoleucine), L-류신(L-Leucine), L-트립토판(L-Tryptophan), L-메티오닌(L-Methionine), 호모세린(Homoserine), N,N-디메틸글라이신(N,N-Dimethylglycine), S-락테이트(S-Lactate), 글리세롤 3-포스페이트(Glycerol 3-phosphate), L-말산(L-Malic acid) 및 하이포잔틴(Hypoxanthine)은 항생제 치료 전의 마이코박테리움 아비움 복합체(MAC) 감염 환자에서의 중앙값 대비 지속적으로 균 양성이 발생한 환자 군에서 유의적으로 그 발현이 증가하였고, 그 외에 알란토인(Allantoin)은 항생제 치료 전의 마이코박테리움 아비움 복합체(MAC) 감염 환자에서의 중앙값 대비 지속적으로 균 양성이 발생한 환자 군에서 유의적으로 그 발현이 감소한 것을 확인할 수 있었다. As shown in Tables 4 and 5 and FIGS. 1 to 13, among the blood metabolites, L-valine, L-threonine, L-isoleucine, and L-leucine. (L-Leucine), L-tryptophan (L-Tryptophan), L-methionine (L-Methionine), homoserine, N,N-dimethylglycine (N,N-Dimethylglycine), S-lactate (S -Lactate), glycerol 3-phosphate, L-malic acid, and hypoxanthine compared with the median value in patients with Mycobacterium avium complex (MAC) infection before antibiotic treatment The expression was significantly increased in the patient group that was continuously positive, and allantoin was continuously positive compared to the median value of patients infected with Mycobacterium avium complex (MAC) before antibiotic treatment. It was confirmed that the expression was significantly reduced in the patient group.
이를 통하여 대사체로, L-발린(L-Valine), L-트레오닌(L-Threonine), L-이소류신(L-Isoleucine), L-류신(L-Leucine), L-트립토판(L-Tryptophan), L-메티오닌(L-Methionine), 호모세린(Homoserine), N,N-디메틸글라이신(N,N-Dimethylglycine), S-락테이트(S-Lactate), 글리세롤 3-포스페이트(Glycerol 3-phosphate), L-말산(L-Malic acid), 하이포잔틴(Hypoxanthine) 및 알란토인(Allantoin)을 비결핵 항산균에 의한 감염 후 항생제 투여 없이는 지속적으로 균 양성이 발생할 것을 예측하여 항생제 치료 여부를 결정하기 위한 바이오마커로 사용할 수 있음을 알 수 있었다.Through this, as metabolites, L-valine, L-threonine, L-isoleucine, L-leucine, L-tryptophan, L-methionine (L-Methionine), homoserine (Homoserine), N,N-dimethylglycine (N,N-Dimethylglycine), S-lactate (S-Lactate), glycerol 3-phosphate (Glycerol 3-phosphate), L-Malic acid, hypoxanthine, and allantoin are biomarkers for determining whether to treat with antibiotics by predicting that bacteria will continue to be positive without antibiotic administration after infection with non-tuberculous mycobacteria. found that it can be used as
본 발명은 비결핵 항산균에 의한 감염 후 예후로, 특히는 적절한 치료 없이는 자발적 균 음전이 발생하지 않고 지속적 균 양성이 발생하는 지 여부를 예측하기 위한 바이오마커와, 상기 예측을 위한 키트 또는 예측 방법에 관한 것이다. The present invention relates to a prognosis after infection by non-tuberculous mycobacteria, in particular, a biomarker for predicting whether or not spontaneous bacterial negative transformation does not occur and persistent bacterial positivity occurs without appropriate treatment, and a kit or prediction method for the prediction is about

Claims (26)

  1. 아미노산(amino acid), 아미노산 유도체, 알란토인(Allantoin), N,N-디메틸글라이신(N,N-Dimethylglycine), 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 후 예후 예측용 바이오마커 조성물.amino acid, amino acid derivative, allantoin, N,N-dimethylglycine, hypoxanthine, lactate, malic acid and glycerol 3-phosphate (Glycerol 3-phosphate) A biomarker composition for predicting prognosis after infection of non-tuberculous mycobacteria, comprising one or more metabolites selected from the group consisting of.
  2. 제1항에 있어서, According to claim 1,
    상기 예후는 비결핵 항산균의 감염 후 치료 없이 균 양성의 지속 여부인 것인, 비결핵 항산균의 감염 후 예후 예측용 바이오마커 조성물.The prognosis is a biomarker composition for predicting the prognosis after infection with non-tuberculous mycobacteria, which is whether the positivity continues without treatment after infection with non-tuberculous mycobacteria.
  3. 제1항에 있어서, According to claim 1,
    상기 예후는 비결핵 항산균의 감염 후 1 개월 이상 균 양성의 발생 여부인 것인, 비결핵 항산균의 감염 후 예후 예측용 바이오마커 조성물.The prognosis is a biomarker composition for predicting prognosis after infection with non-tuberculous mycobacterium, which is whether or not the occurrence of positivity for one month or more after infection with non-tuberculous mycobacterium.
  4. 제1항에 있어서, According to claim 1,
    상기 아미노산 및 아미노산 유도체는 발린(Valine), 트레오닌(Threonine), 이소류신(Isoleucine), 류신(Leucine), 트립토판(Tryptophan), 메티오닌(Methionine) 및 호모세린(Homoserine)으로 이루어진 군에서 선택된 1종 이상을 포함하는, 비결핵 항산균의 감염 후 예후 예측용 바이오마커 조성물.The amino acids and amino acid derivatives include at least one selected from the group consisting of valine, threonine, isoleucine, leucine, tryptophan, methionine, and homoserine. A biomarker composition for predicting prognosis after infection with non-tuberculous mycobacteria, comprising.
  5. 제1항에 있어서, According to claim 1,
    상기 아미노산은 L-형태(L-form)인, 비결핵 항산균의 감염 후 예후 예측용 바이오마커 조성물.The amino acid is L-form (L-form), a biomarker composition for predicting prognosis after infection of non-tuberculous mycobacteria.
  6. 제1항에 있어서, According to claim 1,
    상기 대사체는 목적하는 개체의 전혈(whole blood), 혈장(plasma) 또는 혈청(serum) 유래인 것인, 비결핵 항산균의 감염 후 예후 예측용 바이오마커 조성물.Wherein the metabolite is derived from whole blood, plasma or serum of a target individual, a biomarker composition for predicting prognosis after infection with non-tuberculous mycobacteria.
  7. 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 후 예후 예측용 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 Selected from the group consisting of Mycobacterium xenopi (M. xenopi), a biomarker composition for predicting prognosis after infection with non-tuberculous mycobacteria.
  8. 아미노산(amino acid), 아미노산 유도체, 알란토인(Allantoin), N,N-디메틸글라이신(N,N-Dimethylglycine), 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 후 예후 예측용 키트.amino acid, amino acid derivative, allantoin, N,N-dimethylglycine, hypoxanthine, lactate, malic acid and glycerol 3-phosphate (Glycerol 3-phosphate) A kit for predicting prognosis after infection with non-tuberculous mycobacteria, comprising a quantitative device for measuring the concentration of one or more metabolites selected from the group consisting of.
  9. 제8항에 있어서, 9. The method of claim 8,
    상기 예후는 비결핵 항산균의 감염 후 치료 없이 균 양성의 지속 여부인 것인, 비결핵 항산균의 감염 후 예후 예측용 키트.The prognosis kit for predicting the prognosis after infection with non-tuberculous mycobacterium, the prognosis will be whether the positivity continues without treatment after infection with non-tuberculous mycobacteria.
  10. 제8항에 있어서, 9. The method of claim 8,
    상기 예후는 비결핵 항산균의 감염 후 1 개월 이상 균 양성의 발생 여부인 것인, 비결핵 항산균의 감염 후 예후 예측용 키트.The prognosis kit for predicting the prognosis after infection with non-tuberculous mycobacterium, the prognosis will be whether or not the occurrence of positive bacteria for more than 1 month after infection with non-tuberculous mycobacterium.
  11. 제8항에 있어서, 9. The method of claim 8,
    상기 아미노산 및 아미노산 유도체는 발린(Valine), 트레오닌(Threonine), 이소류신(Isoleucine), 류신(Leucine), 트립토판(Tryptophan), 메티오닌(Methionine) 및 호모세린(Homoserine)으로 이루어진 군에서 선택된 1종 이상을 포함하는, 비결핵 항산균의 감염 후 예후 예측용 키트.The amino acids and amino acid derivatives include at least one selected from the group consisting of valine, threonine, isoleucine, leucine, tryptophan, methionine, and homoserine. A kit for predicting prognosis after infection with non-tuberculous mycobacteria, including.
  12. 제8항에 있어서, 9. The method of claim 8,
    상기 아미노산은 L-형태(L-form)인, 비결핵 항산균의 감염 후 예후 예측용 키트.The amino acid is L-form (L-form), a kit for predicting prognosis after infection of non-tuberculous mycobacteria.
  13. 제8항에 있어서, 9. The method of claim 8,
    상기 대사체는 목적하는 개체의 전혈(whole blood), 혈장(plasma) 또는 혈청(serum) 유래인 것인, 비결핵 항산균의 감염 후 예후 예측용 키트.Wherein the metabolite is derived from whole blood, plasma or serum of a target individual, a kit for predicting prognosis after infection with non-tuberculous mycobacteria.
  14. 제8항에 있어서, 9. The method of claim 8,
    상기 정량 장치는 핵자기 공명 분광 분석기 (NMR), 크로마토그래피 또는 질량분석기인, 비결핵 항산균의 감염 후 예후 예측용 키트.The quantitative device is a nuclear magnetic resonance spectrometer (NMR), chromatography or mass spectrometer, a kit for predicting prognosis after infection with non-tuberculous mycobacteria.
  15. 제8항에 있어서,9. The method of claim 8,
    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 후 예후 예측용 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 Selected from the group consisting of Mycobacterium xenopi (M. xenopi), a kit for predicting the prognosis after infection with non-tuberculous mycobacteria.
  16. 목적하는 개체로부터 분리된 생물학적 시료에서 아미노산(amino acid), 아미노산 유도체, 알란토인(Allantoin), N,N-디메틸글라이신(N,N-Dimethylglycine), 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 후 예후를 예측하기 위한 정보 제공 방법.Amino acids, amino acid derivatives, allantoin, N,N-dimethylglycine, hypoxanthine, lactate, malic acid from a biological sample isolated from a subject of interest (Malic acid) and glycerol 3-phosphate (Glycerol 3-phosphate) information providing method for predicting the prognosis after infection of non-tuberculous mycobacteria comprising measuring the expression level of one or more metabolites selected from the group consisting of .
  17. 제16항에 있어서, 17. The method of claim 16,
    상기 예후는 비결핵 항산균의 감염 후 치료 없이 균 양성의 지속 여부인 것인, 비결핵 항산균의 감염 후 예후를 예측하기 위한 정보 제공 방법.The method for providing information for predicting the prognosis after infection with non-tuberculous mycobacteria, wherein the prognosis is whether or not the positivity continues without treatment after infection with non-tuberculous mycobacteria.
  18. 제16항에 있어서, 17. The method of claim 16,
    상기 예후는 비결핵 항산균의 감염 후 1 개월 이상 균 양성의 발생 여부인 것인, 비결핵 항산균의 감염 후 예후를 예측하기 위한 정보 제공 방법.The method for providing information for predicting the prognosis after infection with non-tuberculous mycobacteria, wherein the prognosis is whether or not the occurrence of positive bacteria for at least 1 month after infection with non-tuberculous mycobacteria.
  19. 제16항에 있어서, 17. The method of claim 16,
    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 후 예후를 예측하기 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 at least one selected from the group consisting of, an information providing method for predicting the prognosis after infection with non-tuberculous mycobacteria.
  20. 제16항에 있어서, 17. The method of claim 16,
    상기 아미노산 및 아미노산 유도체는 발린(Valine), 라이신(Lysine), 트레오닌(Threonine), 이소류신(Isoleucine), 류신(Leucine), 트립토판(Tryptophan), 메티오닌(Methionine) 및 호모세린(Homoserine)으로 이루어진 군에서 선택된 1종 이상을 포함하는, 비결핵 항산균의 감염 후 예후를 예측하기 위한 정보 제공 방법.The amino acids and amino acid derivatives are from the group consisting of valine, lysine, threonine, isoleucine, leucine, tryptophan, methionine and homoserine. A method for providing information for predicting the prognosis after infection with non-tuberculous mycobacterium, comprising at least one selected.
  21. 제16항에 있어서, 17. The method of claim 16,
    상기 아미노산은 L-형태(L-form)인, 비결핵 항산균의 감염 후 예후를 예측하기 위한 정보 제공 방법.The amino acid is an L-form (L-form), information providing method for predicting the prognosis after infection of non-tuberculous mycobacteria.
  22. 제16항에 있어서, 17. The method of claim 16,
    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 후 예후를 예측하기 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. A method of providing information for predicting the prognosis.
  23. 제16항에 있어서, 17. The method of claim 16,
    상기 대사체의 발현 수준을 측정하는 단계는 핵자기 공명 분광 분석기 (NMR), 크로마토그래피 또는 질량분석기인 정량 장치를 이용하여 수행되는, 비결핵 항산균의 감염 후 예후를 예측하기 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 predicting the prognosis after infection of non-tuberculous mycobacteria.
  24. 제20항에 있어서,21. The method of claim 20,
    상기 목적하는 개체의 생물학적 시료에 대하여 측정된 발린(Valine), 트레오닌(Threonine), 이소류신(Isoleucine), 류신(Leucine), 트립토판(Tryptophan), 메티오닌(Methionine), 호모세린(Homoserine), N,N-디메틸글라이신(N,N-Dimethylglycine), 하이포잔틴(Hypoxanthine), 락테이트(Lactate), 말산(Malic acid) 및 글리세롤 3-포스페이트(Glycerol 3-phosphate)로 이루어진 군에서 선택된 1종 이상의 발현 수준이 대조군에 비하여 증가된 경우, 비결핵 항산균에 의한 감염 후 치료 없이는 균 양성이 지속되거나 지속될 가능성이 높은 것으로 예측하는 단계를 더 포함하는, 비결핵 항산균의 감염 후 예후를 예측하기 위한 정보 제공 방법.Valine, Threonine, Isoleucine, Leucine, Tryptophan, Methionine, Homoserine, N,N measured with respect to the biological sample of the subject of interest -Dimethyl glycine (N,N-Dimethylglycine), hypoxanthine (Hypoxanthine), lactate (Lactate), malic acid (Malic acid) and glycerol 3-phosphate (Glycerol 3-phosphate) at least one expression level selected from the group consisting of When it is increased compared to the control group, the method of providing information for predicting the prognosis after infection with non-tuberculous mycobacterium, further comprising the step of predicting that the positivity continues or is highly likely to continue without treatment after infection with non-tuberculous mycobacteria .
  25. 제20항에 있어서,21. The method of claim 20,
    상기 목적하는 개체의 생물학적 시료에 대하여 측정된 알란토인(Allantoin)의 발현 수준이 대조군에 비하여 감소된 경우, 비결핵 항산균에 의한 감염 후 치료 없이는 균 양성이 지속되거나 지속될 가능성이 높은 것으로 예측하는 단계를 더 포함하는, 비결핵 항산균의 감염 후 예후를 예측하기 위한 정보 제공 방법.If the expression level of allantoin measured with respect to the biological sample of the target individual is reduced compared to the control, the bacterial positivity continues or is highly likely to continue without treatment after infection with non-tuberculous mycobacteria. The step of predicting Further comprising, a method for providing information for predicting the prognosis after infection with non-tuberculous mycobacteria.
  26. 제16항에 있어서,17. The method of claim 16,
    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 후 예후를 예측하기 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 predicting the prognosis after infection of non-tuberculous mycobacteria.
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