CN113252806B - Use of S-adenosyl homocysteine in the preparation of a product for the diagnosis or treatment of Kawasaki disease - Google Patents
Use of S-adenosyl homocysteine in the preparation of a product for the diagnosis or treatment of Kawasaki disease Download PDFInfo
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- CN113252806B CN113252806B CN202110321840.8A CN202110321840A CN113252806B CN 113252806 B CN113252806 B CN 113252806B CN 202110321840 A CN202110321840 A CN 202110321840A CN 113252806 B CN113252806 B CN 113252806B
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- 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
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- 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
- G01N30/04—Preparation or injection of sample to be analysed
- G01N30/06—Preparation
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- 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
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- 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
- G01N30/86—Signal analysis
- G01N30/8624—Detection of slopes or peaks; baseline correction
- G01N30/8631—Peaks
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- 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
- G01N30/04—Preparation or injection of sample to be analysed
- G01N2030/042—Standards
- G01N2030/045—Standards internal
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- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
- Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change
Abstract
The application discloses application of S-adenosyl homocysteine in preparation of a product for diagnosing or treating Kawasaki disease, which screens, identifies and finally confirms that the S-adenosyl homocysteine in serum can be used as a biomarker for early diagnosis of the Kawasaki disease through metabonomics for the first time, and develops a diagnosis product for early diagnosis of the Kawasaki disease based on the biomarker. Meanwhile, the kit has the characteristics of low detection cost and good repeatability, and the stability and reliability of the S-adenosyl homocysteine in serum for diagnosing Kawasaki disease are verified in a discovery and verification mode, so that the kit has important clinical development and application values.
Description
Technical Field
The application relates to the technical field of medical biological detection, in particular to application of S-adenosyl homocysteine as a biomarker or a drug target in preparation of a product for diagnosing or treating Kawasaki disease.
Background
Kawasaki disease is an acute self-limiting systemic vasculitis which is well developed in children under the age of 5 years, and the main complication is coronary artery injury, and 20-25% of Kawasaki patients can form coronary tumors and even myocardial infarction under the condition of not being treated in time. In recent years, the incidence of Kawasaki disease increases year by year, and currently exceeds rheumatic fever, and is one of the important factors causing infantile acquired heart disease in developed countries and developing countries. The pathogenesis of Kawasaki disease is not clear, and the diagnosis mode is mainly based on clinical manifestations (such as fever, waxberry tongue, hand and foot molting, rash, etc. lasting more than 5 days). However, since the main symptoms of kawasaki disease are not specific and often do not occur simultaneously, many febrile diseases such as respiratory tract infection, drug eruption, scarlet fever, epstein barr virus infection, measles, infant idiopathic arthritis, etc. and kawasaki disease have similar clinical manifestations, bring about difficulty in diagnosis and often cause misdiagnosis or omission. Therefore, in order to improve the sensitivity and specificity of early clinical diagnosis of kawasaki disease, development of new diagnostic products is imperative.
Disclosure of Invention
The application discovers a biomarker S-adenosyl homocysteine for early diagnosis of Kawasaki disease, and develops a diagnosis product for early diagnosis of the Kawasaki disease based on the biomarker, and the diagnosis product can solve the problems of difficult early diagnosis and easy misdiagnosis of the Kawasaki disease.
Based on the discovery that S-adenosyl homocysteine can be used as a biomarker for early diagnosis of Kawasaki disease:
the application provides an application of S-adenosyl homocysteine as a diagnosis marker of Kawasaki disease in preparing a diagnosis product of Kawasaki disease.
Optionally, the test sample of the diagnostic product is serum of the subject.
Optionally, the diagnostic product is a diagnostic kit, and the diagnostic kit comprises a detection reagent for specifically detecting S-adenosyl homocysteine in a biological sample.
The application also provides application of the reagent for in-vitro detection of S-adenosyl homocysteine in serum in preparation of Kawasaki disease diagnosis products. Alternatively, the diagnostic product is a diagnostic kit or a detection probe.
The application also provides an application of the S-adenosyl homocysteine serving as a medicine target spot of Kawasaki disease in preparing medicines for treating the Kawasaki disease.
The application also provides a Kawasaki disease diagnosis kit based on human serum S-adenosyl homocysteine detection, which comprises a detection reagent for specifically detecting the S-adenosyl homocysteine in the serum of a subject.
Optionally, the kawasaki disease diagnosis kit comprises:
(1) S-adenosyl homocysteine standard;
(2) A dilution liquid;
(3) A derivatizing agent;
(4) An internal standard solution;
(5) Eluting the eluent;
(6) Protein solutions mimicking the serum environment.
Optionally, the diluent is an acetonitrile solution containing formic acid; the derivatization reagent is triethylamine, phenyl isothiocyanate or phthalic dicarboxaldehyde; the internal standard solution is an acetonitrile solution of DL-2 amino pimelic acid; the mobile phase A of the eluent is an ammonium formate solution containing formic acid, and the mobile phase B is acetonitrile; the protein solution is bovine serum albumin solution.
Further, the mass percentage content of formic acid in the acetonitrile solution containing formic acid is 0.05-0.15%, and acetonitrile is acetonitrile solution with mass percentage of 90%; in the acetonitrile solution of DL-2 amino pimelic acid, the concentration of the DL-2 amino pimelic acid is 55.0 mug/mL, and acetonitrile is acetonitrile solution with the mass fraction of 80.0%; in the ammonium formate solution containing formic acid, the mass percentage of formic acid is 0.005-0.02%; the concentration of the bovine serum albumin in the bovine serum albumin solution is 45.0 mg/mL-50.0 mg/mL.
Most preferably, the diluent is a 90% acetonitrile solution containing 0.1% formic acid; the derivatization reagent is triethylamine, phenyl isothiocyanate or phthalic dicarboxaldehyde; the internal standard solution is an 80.0% acetonitrile solution of DL-2 amino pimelic acid, wherein the concentration of the DL-2 amino pimelic acid is 55.0 mug/mL; the mobile phase A of the eluent is ammonium formate solution containing 0.01% formic acid, and the mobile phase B is acetonitrile (pure acetonitrile); the protein solution is a bovine serum albumin solution with the concentration of 50.0mg/mL.
The application also provides a detection system of S-adenosyl homocysteine in serum, which comprises the following steps:
the standard curve drawing module is used for drawing a quantitative standard curve based on the fact that S-adenosyl homocysteine is used as a standard substance, DL-2 amino pimelic acid is used as an internal standard, the ratio of peak areas of the S-adenosyl homocysteine and the DL-2 amino pimelic acid is used as a vertical axis, and the concentration of the S-adenosyl homocysteine is used as a horizontal axis;
a separation module for extracting and eluting S-adenosyl homocysteine from a serum sample from a subject and recording the peak area thereof;
and the calculation module is used for calculating the concentration of the S-adenosyl homocysteine in the serum sample of the subject based on the standard curve drawn by the standard curve drawing module and the peak area recorded by the separation module.
Optionally, the separation module comprises an HILIC chromatographic column and an ultra-high performance liquid chromatograph tandem mass spectrometer.
Compared with the prior art, the application has the following effects:
the application screens, identifies and finally confirms the S-adenosyl homocysteine in serum through metabonomics for the first time, can be used as a biomarker for early diagnosis of Kawasaki disease, is used for diagnosis of the Kawasaki disease, has the characteristics of high sensitivity and high specificity, and simultaneously has the characteristics of low detection cost and good repeatability, and the stability and the reliability of the S-adenosyl homocysteine in serum for diagnosis of the Kawasaki disease are verified through a discovery and verification mode, so that the application has important clinical development and application values.
Drawings
FIG. 1 is a graph of the change in S-adenosyl homocysteine content (mean.+ -. Standard error) in serum samples of Kawasaki patients, kawasaki disease-complicated coronary lesions, non-Kawasaki disease-complicated coronary lesions, FC-complicated lesions, HC-complicated control subjects, CKD-complicated patients, and cured Kawasaki patients (KD-nonCALS-Kawasaki disease; KD-CALS-Kawasaki disease-complicated coronary lesions).
FIG. 2 is a graph of ROC of S-adenosyl homocysteine for patients with Kawasaki disease and for patients with non-Kawasaki disease fever; wherein A shows the ROC curve of S-adenosyl homocysteine in the experimental group for patients with Kawasaki disease and patients with non-Kawasaki disease fever; b shows ROC curves verifying the use of S-adenosyl homocysteine in patients with kawasaki disease and those with non-kawasaki disease fever.
Detailed Description
The following description of the embodiments of the present application will be made clearly and completely with reference to the accompanying drawings, in which it is apparent that the embodiments described are only some embodiments of the present application, but not all embodiments. All other embodiments, which can be made by those skilled in the art based on the embodiments of the application without making any inventive effort, are intended to be within the scope of the application.
Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this application belongs. The terminology used herein in the description of the application is for the purpose of describing particular embodiments only and is not intended to be limiting of the application.
Metabonomics is an emerging discipline after metagenomics, proteomics and transcriptomics, a scientific field aimed at studying small molecule metabolites. Because the presence of small molecule metabolites in body fluids is relatively stable, analysis of some endogenous small molecule metabolites by metabolomics has been widely used for early diagnosis of disease. The application detects the metabolite in serum by using a metabonomics method combined with ultra-high performance liquid chromatography-mass spectrometry, and screens out the early diagnosis biomarker suitable for Kawasaki disease through bioinformatics analysis. The method has the advantages of quick detection, good repeatability, high sensitivity and low cost. The potential diagnosis markers are screened by adopting a strategy from discovery to verification, the Kawasaki disease early diagnosis markers based on human serum S-adenosyl homocysteine are successfully screened, the diagnosis sensitivity and the specificity of the diagnosis markers are good, and the diagnosis markers are not reported in the kit.
The application discovers that S-adenosyl homocysteine can be used as a biomarker for early diagnosis of Kawasaki disease, and the process is as follows:
(1) The metabonomics fingerprint analysis is carried out on the serum of the Kawasaki patient, the non-Kawasaki fever patient and the healthy control subject by utilizing the metabonomics technology of the ultra-high performance liquid chromatograph tandem mass spectrometer, the change of choline metabolism and amino acid metabolism channels in the serum of the Kawasaki patient is found, and the remarkable difference of 17 metabolites between the serum of the Kawasaki patient and the serum of the non-Kawasaki fever patient is found.
(2) The 17 metabolites were subjected to ROC analysis (Receiver Operating Characteristic Curve) using data statistics software SPSS, and metabolites optimal for kawasaki disease diagnosis were screened out and evaluated for sensitivity and specificity. The results are shown in FIG. 2A, which shows the ROC curve of the S-adenosyl homocysteine in the experimental group for the differential diagnosis of patients suffering from Kawasaki disease and patients suffering from non-Kawasaki disease, and finally, the S-adenosyl homocysteine has the best performance for diagnosing the Kawasaki disease, the AUROC is 0.922, the sensitivity and the specificity are 96.40 percent and 79.10 percent respectively when the cut-off value is 13.98ng/mL, namely, the S-adenosyl homocysteine content in serum of a febrile infant is lower than 13.98ng/mL, and the Kawasaki disease can be diagnosed.
(3) The diagnostic markers found in step (2) were validated using a single set of double-blind experimental samples (including kawasaki patients and non-kawasaki febrile patients), and the results are shown in fig. 2B, which shows ROC curves of the validation set of S-adenosyl homocysteine for differential diagnosis of kawasaki patients and non-kawasaki febrile patients, and S-adenosyl homocysteine was found to be a diagnostic marker for kawasaki disease.
(4) The diagnosis marker found in the step (2) is verified again by using a batch of serum of the patient with the recovered Kawasaki disease, and as a result, the serum S-adenosyl homocysteine content of the patient with the Kawasaki disease is found to be reduced along with the aggravation of the disease, but the serum S-adenosyl homocysteine level returns to the normal level after the treatment, and the serum S-adenosyl homocysteine level is further determined to be used for diagnosing the Kawasaki disease and possibly judging the prognosis of the Kawasaki disease.
Based on the finding:
the present application provides: application of S-adenosyl homocysteine as a diagnosis marker of Kawasaki disease in preparing a diagnosis product of Kawasaki disease; application of reagent for in vitro detection of S-adenosyl homocysteine in serum in preparation of Kawasaki disease diagnosis product; an application of S-adenosyl homocysteine as a medicine target point of Kawasaki disease in preparing medicines for treating the Kawasaki disease; a kawasaki disease diagnostic kit; and a detection system for S-adenosyl homocysteine in serum.
The diagnostic product can be a reagent, test paper, a kit and the like, and optionally, the diagnostic product is a diagnostic kit, and the diagnostic kit comprises a detection reagent for specifically detecting S-adenosyl homocysteine in a biological sample. The test sample of the diagnostic product is the serum of the subject.
As one embodiment of the kawasaki disease diagnostic kit, the kawasaki disease diagnostic kit comprises:
(1) S-adenosyl homocysteine standard;
(2) A dilution liquid;
(3) A derivatizing agent;
(4) An internal standard solution;
(5) Eluting the eluent;
(6) Protein solutions mimicking the serum environment.
The S-adenosyl homocysteine standard is used for qualitative analysis of S-adenosyl homocysteine in serum and drawing of a marking curve. The mass numbers of parent ions and main fragment ions of S-adenosyl homocysteine in the MRM mode of mass spectrum are 385.08 and 136.055 respectively.
The dilutions are used to pre-treat the extract and standard of a serum sample from a subject, optionally in 90% acetonitrile containing 0.1% formic acid.
Derivatizing agents include triethylamine, phenyl isothiocyanate, or o-phthalaldehyde solutions, and the like.
The internal standard solution can be selected from 80.0% acetonitrile solution of DL-2 aminopimelic acid, the concentration of DL-2 aminopimelic acid is 55.0 mug/mL, quantitative analysis is carried out in a mass spectrum SRM or MRM mode, and the mass numbers of parent ions and main fragment ions thereof in the mass spectrum MRM mode are 176.10 and 112.10 respectively.
The eluent is used for eluting the chromatographic column, alternatively, mobile phase A can be selected from ammonium formate solution containing 0.01% formic acid, and mobile phase B can be selected from acetonitrile. The chromatographic column can be C18 or HILIC chromatographic column.
The protein solution mimicking the serum environment may be selected as a bovine serum albumin solution having a concentration of 50.0mg/mL.
The kit can be applied to an ultra-high performance liquid chromatograph tandem mass spectrometer, can also detect more than 20 amino acids or metabolites related to one-carbon metabolism at the same time, and can more comprehensively analyze metabolic abnormalities of Kawasaki patients. Serum samples were used to test the effect of the application. The serum samples of the Kawasaki disease patients and the non-Kawasaki disease fever patients are adopted, and the serum S-adenosyl homocysteine is used as a diagnosis standard, so that the two groups of patients can be well distinguished.
Specifically, a method for detecting S-adenosyl homocysteine in a serum sample from a subject using a kit of the application comprises the steps of:
(1) Using S-adenosyl homocysteine as a standard substance, adding an internal standard DL-2 amino pimelic acid, carrying out derivatization or direct sample injection analysis, and drawing a corresponding quantitative standard curve;
(2) Serum sample pretreatment from subjects: thawing a serum sample at 4 ℃, adding 400.0 mu L of 90.0% acetonitrile solution containing 0.1% formic acid and 50.0 mu L of internal standard solution into 50.0 mu L of serum, vigorously vortex and mix for 15.0S, centrifuging 14000g for 10.0min, taking 200.0 mu L of supernatant in an automatic sample injection bottle for detection, and recording the ratio of the eluting peak area to the internal standard peak area of S-adenosyl homocysteine in the serum sample;
(3) Calculating the content of S-adenosyl homocysteine in the serum sample of the subject according to the ratio of the quantitative standard curve of the step (1) to the peak area of the step (2).
The diagnosis kit detects the content of S-adenosyl homocysteine in serum of febrile patients (including patients suffering from Kawasaki disease and non-Kawasaki disease), takes 13.98ng/mL as a cut-off value, and diagnoses the Kawasaki disease when the content of S-adenosyl homocysteine in the serum of the patients is lower than 13.98 ng/mL.
One embodiment of a system for detecting S-adenosyl homocysteine in serum comprises:
the standard curve drawing module is used for drawing a quantitative standard curve based on the fact that S-adenosyl homocysteine is used as a standard substance, DL-2 amino pimelic acid is used as an internal standard, the ratio of peak areas of the S-adenosyl homocysteine and the DL-2 amino pimelic acid is used as a vertical axis, and the concentration of the S-adenosyl homocysteine is used as a horizontal axis;
a separation module for extracting and eluting S-adenosyl homocysteine from a serum sample from a subject and recording the peak area thereof;
and the calculation module is used for calculating the concentration of the S-adenosyl homocysteine in the serum sample of the subject based on the standard curve drawn by the standard curve drawing module and the peak area recorded by the separation module.
A diagnostic system comprising: the chromatographic column is HILIC chromatographic column (2.1X105 mm,3.5 μm, merck, US), the detecting instrument is ultra-high performance liquid chromatography tandem mass spectrometer, the metabolite is detected under multiple reaction monitoring mode (MRM), and the electrospray ion source adopts positive ions.
One specific embodiment of the kawasaki disease diagnostic kit comprises:
(1) S-adenosyl homocysteine standard;
(2) Dilution liquid: a 90% acetonitrile solution containing 0.1% formic acid;
(3) Derivatizing agent: derivatizing agents include triethylamine, phenyl isothiocyanate, or o-phthalaldehyde solutions, and the like.
(4) Internal standard solution: DL-2 amino pimelic acid in 80.0% acetonitrile at a concentration of 55.0. Mu.g/mL;
(5) Eluent: mobile phase A is ammonium formate solution containing 0.01% formic acid, mobile phase B is acetonitrile;
(6) Protein solution mimicking the serum environment: a bovine serum albumin solution at a concentration of 50.0mg/mL.
Specific examples of the detection using the kit are described below, and the concentrations defined by percentages in the examples below are mass percentages unless otherwise specified.
Example 1:
1. serum sample collection
Prior to collection, all subjects who were included in the study were taken with signed informed consent.
50 febrile patients (including 30 patients with Kawasaki disease and 20 patients with non-Kawasaki disease) all fasted for more than 8 hours and were venipuncture blood taken the next morning. After whole blood is collected, standing for 1 hour, centrifuging for 10 minutes at 2000g, and collecting upper serum and storing in a refrigerator at-80 ℃.
2. Analysis method
2.1 drawing of a Standard Curve
10.0mg of S-adenosyl homocysteine standard is precisely weighed, and diluted into different concentration gradients by taking 90% acetonitrile solution containing 0.1% formic acid as a solvent. 50.0 mu L of protein solution (bovine serum albumin solution with the concentration of 50.0 mg/mL) is taken, 50.0 mu L of internal standard solution, 50.0 mu L of standard solution and 350.0 mu L of 90% acetonitrile solution containing 0.1% formic acid are sequentially added, the mixture is vigorously vortexed and mixed for 15.0s, 14000g is centrifuged for 10.0min, and 200.0 mu L of supernatant is taken in an automatic sample injection bottle for drawing a standard curve.
2.2 pretreatment of serum samples
Thawing a serum sample at 4 ℃, adding 400.0 mu L of 90.0% acetonitrile solution containing 0.1% formic acid and 50.0 mu L of internal standard solution into 50.0 mu L of serum, vigorously vortex mixing for 15.0s, centrifuging for 10.0min with 14000g, taking 200.0 mu L of supernatant, derivatizing the phthalic aldehyde solution, and using in an automatic sample injection bottle for detection.
2.3 analysis by ultra high Performance liquid chromatography tandem Mass Spectrometry
(1) Liquid phase conditions: the chromatograph is Waters ultra-high performance liquid chromatography; the chromatographic column is HILIC chromatographic column; mobile phase A is ammonium formate solution containing 0.01% formic acid, mobile phase B is acetonitrile; the flow rate is 0.5mL/min; the sample injection volume is 5.0 mu L; the separation time is 12.0min; elution was performed using the following gradient: 0-2.5min,20% A;2.5-5.0min,20% -80% A;5.0-8.0min,80% A;8-8.5min,80% -20% A;8.5-12.0min,20% A. The column temperature was set at 45 ℃ and the autosampler temperature was set at 8 ℃.
(2) Mass spectrometry conditions: the mass spectrometer is Xex QTof (Waters, US), the metabolites are detected in a multiple reaction monitoring mode (MRM), and the electrospray ion source adopts positive ion detection; the electrospray capillary voltage was set at 3.0kV, nitrogen was used as the drying gas for solvent evaporation, and the flow rate was 50L/h; the ion source temperature was 100 ℃.
2.4 serum detection results
Drawing a standard curve by taking the concentration of the standard substance as an abscissa and the ratio of the peak area of the standard substance to the peak area of the internal standard substance as an ordinate; substituting the detection result of the S-adenosyl homocysteine in the serum sample into a standard curve to finally obtain the content of the S-adenosyl homocysteine in each sample.
As shown in FIG. 1, the results show that the content of S-adenosyl homocysteine in serum of Kawasaki patients is greatly reduced relative to that of non-Kawasaki fever patients, and the accuracy of distinguishing two groups of patients is 86.67% and the false positive rate is 10.00% when 13.98ng/mL is taken as a cut-off value.
The above examples illustrate only a few embodiments of the application, which are described in detail and are not to be construed as limiting the scope of the application. It should be noted that it will be apparent to those skilled in the art that several variations and modifications can be made without departing from the spirit of the application, which are all within the scope of the application. Accordingly, the scope of protection of the present application is to be determined by the appended claims.
Claims (6)
- Use of S-adenosyl homocysteine as a diagnostic marker for kawasaki disease in the manufacture of a product for diagnosis of kawasaki disease, which is diagnosed as kawasaki disease when the S-adenosyl homocysteine content in the serum of a subject is lower than 13.98 ng/mL.
- 2. The use according to claim 1, wherein the test sample of the diagnostic product is serum of a subject.
- 3. The use according to claim 1, wherein the diagnostic product is a diagnostic kit comprising a detection reagent for specifically detecting S-adenosylhomocysteine in a biological sample.
- 4. Use of a reagent for in vitro detection of S-adenosyl homocysteine in serum in the preparation of a kawasaki disease diagnostic product, for diagnosing kawasaki disease when the S-adenosyl homocysteine content in the serum of a subject is below 13.98 ng/mL.
- 5. The use according to claim 4, wherein the diagnostic product is a diagnostic kit or a detection probe.
- The application of 6.S-adenosyl homocysteine as a drug target point of Kawasaki disease in preparing a drug for treating the Kawasaki disease is that the Kawasaki disease is diagnosed when the S-adenosyl homocysteine content in serum of a subject is lower than 13.98 ng/mL.
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