Diagnostic method for behcet's disease using metabolome analysis
A technology for Behcet's disease and metabolites, applied in the field of diagnosis of Behcet's disease, can solve the problems of no diagnosis and prediction of Behcet's disease biomarker research reports, etc., to achieve rapid recovery of daily life, rapid treatment, and reduce long-term Effect
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example 1
[0061] Example 1: Identifying Metabolites Using GC / TOF MS
[0062] 20 microliters of blood each from the Behcet's disease patient group and healthy controls were mixed with 980 microliters of pure methanol, and the mixture was centrifuged to extract metabolites.
[0063] The derivatization process for GC / TOF MS is as follows.
[0064] Each extracted sample was dried in a speed bag, and then 5 μl of 40% (w / v) O-methylhydroxylamine hydrochloride in pyridine was added to it. ), so that they reacted for 90 minutes at 30° C. and 200 revolutions per minute (rpm). Then, 45 microliters of N-methyl-N-(trimethylsilyl)trifluoroacetamide (N-methyl-N-(trimethylsilyl)trifluoroacetamide) was added thereto so that they were heated at 37°C and React for 30 minutes at 200 rpm.
[0065] The instrument conditions of GC / TOF MS are as follows.
[0066] The column used for analysis is an RTX-5Sil MS capillary column (30 meters in length, 0.25 mm in film thickness and 25 mm in inner diameter), an...
example 2
[0071] Example 2: Differences in Metabolite Profiles in Blood Samples of Behcet's Disease Patients and Healthy Controls Using PLS-DA
[0072] The intensity of each metabolite identified according to Example 1 was divided by the sum of the intensities of all identified metabolites to normalize each metabolite. Subsequently, they were subjected to PLS-DA using SIMCA-P+ (version 12.0).
[0073] Such as figure 1 As shown, it was confirmed that there were significant differences in metabolite profiles in the blood samples of Behcet's disease patients and the blood samples of healthy controls.
[0074] Table 2 shows VIP values and loading values, which represent the degree and direction of influence of the 104 metabolites used in the PLS-DA model on the model.
[0075] [Table 2]
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example 3
[0081] Example 3: Selection of Behcet's Disease Patient-Specific Biomarker Metabolites
[0082] In order to find biomarkers that showed a specific increase or decrease in Behcet's disease patients, VIP values, fold changes, AUC values and p-values affecting the difference in the metabolomic profile of each metabolite obtained from Example 2 were obtained. A VIP value of 1.5 or greater than 1.5, a fold change of 1.2, an AUC value of 0.800 or greater than 0.800, and a p value of less than 0.01 were set as the reference values for each metabolite, and 13 metabolites showed positive effects on diagnosis Behcet's disease applicability (see Table 3). In addition, the absolute intensities of these metabolites were compared by group (see figure 2 ).
[0083] Table 3 below shows the VIP value, AUC value, fold change (fold change) and p value (BD: Behcet's disease patient; control : healthy individuals).
[0084] [table 3]
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[0086] AUC, area under the ROC curve; ...
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