CN114252547A - Application of dimethylglycine as serum marker of fetal congenital heart disease - Google Patents
Application of dimethylglycine as serum marker of fetal congenital heart disease Download PDFInfo
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- 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
- G01N30/88—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
Abstract
The application discloses an application of dimethylglycine as a serum marker of fetal congenital heart disease, the application screens, identifies and finally confirms that dimethylglycine in serum can be used as a biomarker for early diagnosis of fetal congenital heart disease through metabonomics for the first time, and a diagnosis product for early diagnosis of fetal congenital heart disease is developed based on the biomarker, and the diagnosis product has high sensitivity and high specificity and can solve the problem of early missed diagnosis of fetal congenital heart disease. Meanwhile, the kit has the characteristics of low detection cost and good repeatability, and the stability and reliability of the dimethylglycine in the serum for diagnosing the fetal congenital heart disease are verified in a discovery and verification mode, so that the kit has important clinical development and application values.
Description
Technical Field
The invention relates to the technical field of medical biological detection, in particular to application of dimethylglycine as a biomarker in preparing a product for diagnosing or treating fetal congenital heart disease.
Background
Congenital heart disease is the most common congenital abnormality, which is a disease caused by abnormal development of cardiovascular vessels in patients due to genetic factors, environmental factors, or both. Statistically, 8-10 (0.8% -1%) of 1000 live births suffer from CHD, and the incidence rate thereof is higher by 8.3% in premature infants, and these newborns (50% -60%) need to be taken for life with repeated surgical treatment, which imposes a heavy burden on families and society.
At present, ultrasonic examination of a fetus plays a crucial role in prenatal diagnosis of CHD, an operator can clearly observe the heart morphological structure of the fetus and is safe and noninvasive for a pregnant woman and the fetus, but missed diagnosis often occurs clinically due to the influence of various factors such as experience of the operator, fetal position, frequency of an instrument probe and the like. Magnetic Resonance Imaging (MRI) can overcome the defects of small ultrasonic diagnosis visual field and poor soft tissue contrast, and the image quality can not be influenced by factors such as amniotic fluid, bone and maternal obesity and the like. However, there is a time difference between MRI image acquisition and real-time imaging, which cannot be checked in real time and is also susceptible to fetal movement.
The abnormality of the CHD patient is not limited to the heart, and is accompanied by many other abnormalities other than the heart, and also by a change in genetic material including chromosomal aberration, monogenic genetic defect, polygenic genetic defect, and the like, and therefore, genetic examination is also important. The chromosome karyotype analysis is a classical genetic examination technology for diagnosing chromosome number abnormality and microstructure abnormality, but the detection period of the technology is long, the resolution is low, and the technical requirements on operators are high. The chromosome set copy number variation detection (CNV-seq) technology has the advantages of rapidness, accuracy, low operation cost and the like. However, this technique does not detect uniparental disomes and triploids. Chromosome Microarray Analysis (CMA) has the advantages of high throughput and high sensitivity, but can only perform detection analysis on known loci, and is relatively expensive.
Therefore, early prenatal diagnosis is crucial to improve outcomes in both mother and fetus, which helps to provide optimal perinatal and perioperative management, reducing postpartum morbidity and mortality. Therefore, in order to improve the sensitivity and specificity of early clinical diagnosis of fetal congenital heart diseases, the development of new diagnostic products is imperative.
Disclosure of Invention
The application finds a biomarker for early diagnosis of fetal congenital heart disease, namely serum N, N-dimethylglycine, and develops a diagnostic product for early diagnosis of fetal congenital heart disease based on the biomarker, wherein the diagnostic product can solve the problem that early misdiagnosis of fetal congenital heart disease is easy.
Based on the discovery that dimethylglycine can be used as a biomarker for early diagnosis of fetal congenital heart disease:
the application provides an application of dimethylglycine as a diagnosis marker of fetal congenital heart disease in preparation of fetal congenital heart disease diagnosis products.
Optionally, the detection sample of the fetal congenital heart disease diagnostic product is serum of the subject.
Optionally, the fetal congenital heart disease diagnostic product is a diagnostic kit, and the diagnostic kit contains a detection reagent for specifically detecting dimethylglycine in the biological sample.
The application also provides an application of the reagent for in vitro detection of dimethylglycine in serum in preparation of a fetal congenital heart disease diagnosis product.
Optionally, the diagnostic product is a diagnostic kit.
Alternatively, the reagent can be a reagent for detecting the content level of dimethylglycine in serum based on the chromatographic and mass spectrum separation principles.
The application also provides application of dimethylglycine as a drug target of fetal congenital heart disease in preparation of a drug for treating fetal congenital heart disease. The application also provides a fetal congenital heart disease diagnostic kit based on dimethylglycine detection in human serum, which comprises a detection reagent for specifically detecting dimethylglycine in serum of a subject.
Optionally, the kit for diagnosing fetal congenital heart disease comprises:
(1) a dimethylglycine standard;
(2) diluting the solution;
(3) an internal standard solution;
(4) eluting the solution;
optionally, the diluent is an acetonitrile solution; the internal standard solution is an isotope labeled dimethyl glycine solution diluted by ammonium formate or an isotope labeled choline (choline-d9) solution diluted by ammonium formate; the mobile phase A of the eluent is an ammonium formate solution containing formic acid, and the mobile phase B of the eluent is acetonitrile.
Further, the acetonitrile solution of the diluent is an acetonitrile solution with a mass fraction of 80% and an acetonitrile solution with a mass fraction of 100%, the concentration of isotope-labeled dimethylglycine or isotope-labeled choline in the internal standard solution is 200-500ng/ml, and the concentration of ammonium formate is 10 mmol/L; in the mobile phase A, the mass percentage of formic acid is 0.005-0.02%.
Most preferably, the diluent is an acetonitrile solution with the mass fraction of 80% and an acetonitrile solution with the mass fraction of 100%; the internal standard solution is an isotope labeling dimethyl nucleotide solution diluted by ammonium formate, wherein the concentration of the isotope labeling dimethyl nucleotide diluted by the ammonium formate is 500ng/ml, and the concentration of the ammonium formate is 10 mmol/L; the mobile phase A of the eluent is ammonium formate solution containing 0.01 percent formic acid, and the mobile phase B of the eluent is acetonitrile.
The present application further provides a dimethylglycine detection system in serum, comprising:
the standard curve drawing module is used for drawing a quantitative standard curve by taking the ratio of the peak areas of the dimethylglycine and the internal standard as a longitudinal axis and the concentration of the dimethylglycine as a transverse axis based on the fact that the dimethylglycine is used as a standard product and the isotope-labeled dimethylnucleotide or isotope-labeled choline is used as the internal standard;
a separation module for extracting and eluting dimethylglycine 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 dimethylglycine 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 a HILIC chromatographic column and an ultra-high performance liquid chromatograph tandem mass spectrometer.
Compared with the prior art, the method has the following effects:
the application screens, identifies and finally confirms that the dimethylglycine in the serum can be used as a biomarker for early diagnosis of the fetal congenital heart disease through metabonomics for the first time, is used for diagnosis of the fetal congenital heart disease, has high sensitivity and high specificity, has the characteristics of low detection cost and good repeatability, verifies the stability and reliability of the dimethylglycine in the serum for diagnosis of the fetal congenital heart disease through a discovery and verification mode, and has important clinical development and application values.
Drawings
FIG. 1 is the chromatogram-mass spectrum related to 6 kinds of choline metabolism in serum.
In fig. 2, a is a graph showing the content change (mean ± sem) of dimethylglycine in serum samples of pregnant women and pregnant women with healthy fetuses, wherein the serum samples are subjected to ultrasonic diagnosis and follow-up to confirm that the fetus is CHD (in the graph, CHDP indicates a pregnant woman subjected to ultrasonic diagnosis and follow-up to confirm that the fetus is CHD, and ZCP indicates a pregnant woman with a healthy fetus).
In FIG. 2, B is a ROC curve of dimethylglycine used in the pregnant women with diagnosed CHD and healthy fetus confirmed by follow-up.
Detailed Description
The technical solutions in the embodiments of the present application will be clearly and completely described below with reference to the drawings in the embodiments of the present application, and it is obvious that the described embodiments are only a part of the embodiments of the present application, and not all of the embodiments. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present 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 present 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 behind phylogenomics, proteomics and transcriptomics, and is a scientific field with the aim of researching small-molecule metabolites. Since the presence of small molecule metabolites in body fluids is relatively stable, the analysis of some endogenous small molecule metabolites through metabolomics has been widely used for early diagnosis of diseases. The method detects the metabolites in the serum by a metabonomics method of ultra-high performance liquid chromatography-mass spectrometry, and screens out the early diagnosis biomarker suitable for fetal congenital heart disease through bioinformatics analysis. The method has the advantages of fast detection, good repeatability, high sensitivity and low cost. The potential diagnostic markers are screened by adopting a strategy from discovery to verification, the early diagnostic marker of fetal congenital heart disease based on human serum dimethylglycine is successfully screened, the diagnostic sensitivity and specificity of the diagnostic marker are good, and a kit of the diagnostic marker is not reported.
The application finds that the process of dimethylglycine as a biomarker for early diagnosis of fetal congenital heart disease is as follows:
(1) the serum of 55 pregnant women (CHDP group) which are subjected to ultrasonic diagnosis and follow-up fetus is CHD and the serum of 49 pregnant women (ZCP group) which are healthy fetuses are subjected to metabonomic fingerprint analysis by utilizing a metabonomic technology based on nuclear magnetic resonance and an ultra-high performance liquid chromatograph tandem mass spectrometer, the change of a choline metabolic pathway in the serum of a patient in the CHDP group is found, and 6 choline related metabolites in the serum of the patient in the CHDP group and the patient in the ZCP group are quantitatively analyzed (see figure 1), and 5 choline metabolites are found to have significant difference.
(2) The 5 metabolites were analyzed by ROC (receptor Operating characteristics Curve) using data statistics software SPSS, and the metabolites with the best diagnostic performance for fetal congenital heart disease were selected and evaluated for sensitivity and specificity. The results are shown in fig. 2, which shows ROC curves of dimethylglycine used for differential diagnosis of ZCP group patients and CHDP group patients in the experimental group, and finally determines that dimethylglycine has the best diagnostic performance for diagnosing fetal congenital heart disease, with AUROC of 0.883, and sensitivity and specificity of 85.2% and 82.0% respectively when the cutoff value is 1.36ug/mL, i.e. when the content of dimethylglycine in serum of patients is higher than 1.36ug/mL, fetal congenital heart disease can be diagnosed.
Based on this finding:
the present application provides: application of dimethylglycine as a diagnostic marker of fetal congenital heart disease in preparation of fetal congenital heart disease diagnostic products; the application of a reagent for in vitro detection of dimethylglycine in serum in the preparation of a fetal congenital heart disease diagnosis product; application of dimethylglycine as a drug target of fetal congenital heart disease in preparing a drug for treating fetal congenital heart disease; a kit for diagnosing fetal congenital heart disease; and a detection system for dimethylglycine in serum.
The diagnostic product can be a reagent, test paper, a kit and the like, and the diagnostic product is a diagnostic kit which contains a detection reagent for specifically detecting dimethylglycine in the biological sample. The test sample of the diagnostic product is the serum of the subject.
As one embodiment of the fetal congenital heart disease diagnostic kit, the fetal congenital heart disease diagnostic kit comprises:
(1) a dimethylglycine standard;
(2) diluting the solution;
(3) an internal standard solution;
(4) eluent
The dimethylglycine standard was used for qualitative analysis of dimethylglycine in serum and for plotting of a standard curve. The mass numbers of the parent ion and the main fragment ion of dimethylglycine in the mass spectrum MRM mode are 104.10 and 58.07 respectively.
The diluent is used for pre-treating the extract and standard substance of serum sample from a subject, and a pure acetonitrile solution and an 80% acetonitrile solution can be selected.
The internal standard solution can be isotope-labeled dimethyl nucleotide solution diluted by ammonium formate or isotope-labeled choline solution diluted by ammonium formate, the concentration of the ammonium formate in the internal standard solution is 10mmol/L, the final concentration of the isotope-labeled dimethyl nucleotide or isotope-labeled choline is 300-500ng/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 of the parent ions in the mass spectrum MRM mode are 113.16 and 69.13 respectively.
The eluent is used for eluting the chromatographic column, and the mobile phase A can be selected from ammonium formate solution containing 0.01% formic acid, and the mobile phase B can be selected from acetonitrile. The chromatographic column can be C18 or HILIC chromatographic column.
The kit can be applied to an ultra-high performance liquid chromatograph tandem mass spectrometer, can also detect other choline related metabolites such as choline, carnitine, acetylcholine and dimethylglycine simultaneously, and can analyze the choline metabolism abnormity of pregnant women with fetal congenital heart diseases more comprehensively. Serum samples were used to test the efficacy of the invention. Serum samples of pregnant women suffering from fetal CHD and pregnant women with healthy fetuses are adopted, and dimethylglycine in the serum is used as a diagnostic standard, so that two groups of patients can be well distinguished.
Specifically, one method for detecting dimethylglycine in a serum sample of a subject using the kit of the present application comprises the steps of:
(1) dimethyl glycine is used as a standard substance, the internal standard is isotope labeling dimethyl nucleotide (the concentration of the isotope labeling dimethyl nucleotide is 500ng/ml) diluted by ammonium formate, then acetonitrile solution is added, direct sample injection detection is carried out, and a corresponding quantitative standard curve is drawn;
(2) serum sample pretreatment from subjects: unfreezing a serum sample at room temperature, adding 30 mu L of internal standard solution (isotope labeled dimethyl nucleotide diluted by ammonium formate), 30 mu L of 80% acetonitrile and 340 mu L of acetonitrile into 50 mu L of serum, violently mixing by vortex, centrifuging for 10.0min at 12000g after ultrasonic treatment, putting 200.0 mu L of supernatant into an automatic sample injection bottle for detecting dimethyl glycine in the serum, and recording the ratio of the elution peak area of the dimethyl glycine in the serum sample to the internal standard peak area;
(3) and (3) calculating the content of the dimethylglycine in the serum sample of the subject according to the ratio of the quantitative standard curve in the step (1) to the peak area in the step (2).
The diagnostic kit detects the content of dimethylglycine in the serum of pregnant women (pregnant women with fetal CHD and healthy fetal pregnant women), and diagnoses that the pregnant women suffer from fetal congenital heart disease with a cutoff value of 1.36ug/mL when the content of dimethylglycine in the serum of patients is higher than 1.36 ug/mL.
One embodiment of the detection system for dimethylglycine in serum comprises:
the standard curve drawing module is used for drawing a quantitative standard curve by taking the ratio of peak areas of dimethyl glycine and an isotope labeled internal standard as a vertical axis and the concentration of the dimethyl glycine as a horizontal axis on the basis of taking the dimethyl glycine as a standard substance and the isotope labeled dimethyl nucleotide or isotope labeled choline (choline-d9) as the internal standard;
a separation module for extracting and eluting dimethylglycine 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 dimethylglycine 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.
The diagnostic system comprises means for: metabolites were detected in a multiplex reaction monitoring mode (MRM) using an UPLC chromatograph of Waters, USA, in combination with a Xevo G2-XS QTof Mass spectrometer on a SeQuant ZIC-HILIC hydrophilic column (3.5 μm, 2.1X 150mm) of Merck, USA, and positive ions were used as the electrospray ion source.
The following is a description of specific examples using the kit for detection, and the concentrations in the following examples using percentage limits refer to mass percentage concentrations unless otherwise specified.
Example 1:
1. serum sample collection
Prior to collection, informed consent was obtained from all subjects enrolled in the study.
104 serum samples from the study were obtained from the maternal care facility of Jiaxing city and the maternal care facility of Jiangxi province, including 55 pregnant women with CHD confirmed by ultrasonic diagnosis and follow-up visit (CHDP group) and 49 pregnant women with healthy fetus (ZCP group).
Inclusion criteria were: firstly, single pregnancy; the B-ultrasonic examination data is complete, and the data is confirmed to be CHD through the birth or labor induction of the follow-up fetus; thirdly, the pregnant woman or the family members sign an informed consent; and fourthly, the pregnant woman is 20-40 years old and pregnant in the middle of gestation (20-24 weeks).
Exclusion criteria: firstly, physical examination can not be carried out according to the specified time or follow-up visit can not be matched; the fetus is combined with other parts to be malformed; ③ pregnant women have serious diseases of heart, brain, liver, kidney and other organs; fourthly, the pregnant women have vaginal hemorrhage, acute fetal intrauterine distress, fetal death, double-fetus and multiple-fetus pregnancy and the like; pregnancy complication or complication pregnant woman such as gestational diabetes and gestational hypertension.
In order to eliminate diet interference, all pregnant women collect blood sample on an empty stomach in the morning and stand still in a refrigerator at 4 ℃ for 2 hours, centrifuge the blood sample at 4000r for 15min, collect serum, subpackage and store in the refrigerator at-80 ℃ and unfreeze at room temperature when using. The study was approved by the ethical committee of the women's healthcare institute, Jiaxing (No.2020-5) and informed consent was obtained from the patients.
2. Analytical method
2.1 drawing of Standard Curve
Weighing a certain amount of dimethylglycine, dissolving the dimethylglycine with distilled water to a certain concentration to obtain mother liquor, and storing the mother liquor at 4 ℃ for later use. And sequentially diluting the mother liquor by 2 times with distilled water, and uniformly mixing the diluted mother liquor by vortex to obtain the dimethylglycine standard solution with different gradients. 50 μ L of dimethylglycine standard solutions with different concentrations and 30 μ L of internal standard solution (isotope labeling dimethyl nucleotide solution diluted by ammonium formate, wherein the concentration of isotope labeling dimethyl nucleotide is 500ng/ml, the concentration of ammonium formate is 10mmol/L), 30 μ L of 80% acetonitrile and 340 μ L of acetonitrile are sequentially added into an EP tube, are mixed by vigorous vortex, are centrifuged for 10.0min at 12000g after ultrasonic treatment, and 200.0 μ L of supernatant is taken out to be placed in an automatic sample injection bottle for drawing a standard curve.
2.2 serum sample pretreatment
Unfreezing a serum sample at room temperature, sequentially adding 50 mu L of serum and 30 mu L of internal standard solution (same as 2.1), 30 mu L of 80% acetonitrile and 340 mu L of acetonitrile into an EP tube, violently vortexing to mix and precipitate protein, centrifuging at 12000g for 10.0min after ultrasonic treatment, and taking 200.0 mu L of supernatant into an automatic sample feeding bottle for detecting dimethylglycine in the serum.
2.3 ultra high performance liquid chromatograph tandem mass spectrometer analysis
(1) Liquid phase conditions: the chromatograph is Waters ultra-performance liquid chromatography; SeQuant ZIC-HILIC hydrophilic chromatography column (3.5 μm, 2.1X 150mm) from Merck, USA; the mobile phase composition is as follows: 10.0mmol/L aqueous ammonium formate solution (A) containing 0.01% formic acid-acetonitrile (B); the flow rate is 0.3mL/min, and the sample injection volume is 4.0 muL; the separation time is 8.0 min; elution was carried out with the following gradient: 0-2.0 min, 10% A; 2.0-4.0min, 10% -70% A; 4.0 min-5.0 min, 70% A; 5.0 min-5.5 min, 70% -10% A; 5.5-8 min, 10% A; the column temperature was set at 45 ℃ and the autosampler temperature at 8 ℃.
(2) Mass spectrum conditions: the mass spectrometer is Xevo QTof (Waters, US), the metabolite is detected in a multiple reaction monitoring mode (MRM), and the electrospray ion source employs positive ion detection; the voltage of the electrospray capillary was set to 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 assay results
Taking the concentration of the standard substance as a horizontal coordinate, taking the ratio of the peak area of the standard substance to the peak area of the internal standard substance as a vertical coordinate, and drawing a standard curve; and substituting the detection result of the dimethylglycine in the serum sample into the standard curve to finally obtain the content of the dimethylglycine in each sample. The serum test results are shown in FIG. 1.
As a result, the serum content of dimethylglycine was found to be greatly increased in patients with CHDP compared to those in ZCP group. When the cut-off value was 1.36ug/mL, the sensitivity and specificity were 85.2% and 82.0%, respectively. Namely, when the content of dimethylglycine in serum of a patient is higher than 1.36ug/mL, fetal congenital heart disease can be diagnosed.
The above-mentioned embodiments only express several embodiments of the present application, and the description thereof is more specific and detailed, but not construed as limiting the scope of the invention. It should be noted that, for a person skilled in the art, several variations and modifications can be made without departing from the concept of the present application, which falls within the scope of protection of the present application. Therefore, the protection scope of the present patent shall be subject to the appended claims.
Claims (10)
1. Application of dimethylglycine as a diagnostic marker of fetal congenital heart disease in preparation of fetal congenital heart disease diagnostic products.
2. The use of claim 1, wherein the test sample of the fetal congenital heart disease diagnostic product is serum of a subject.
3. The use according to claim 1, wherein the product for diagnosing fetal congenital heart disease is a diagnostic kit comprising a detection reagent for specifically detecting dimethylglycine in a biological sample.
4. Application of a reagent for in vitro detection of dimethylglycine in serum in preparation of fetal congenital heart disease products.
5. The use according to claim 4, wherein the product for diagnosing fetal congenital heart disease is a diagnostic kit.
6. Application of dimethylglycine as a drug target of fetal congenital heart disease in preparation of drugs for treating fetal congenital heart disease.
7. A diagnostic kit for fetal congenital heart disease, comprising a detection reagent for specifically detecting dimethylglycine in serum of a subject.
8. The fetal congenital heart disease diagnostic kit according to claim 7, comprising:
(1) a dimethylglycine standard;
(2) diluting liquid: acetonitrile solution;
(3) internal standard solution: an isotopically labeled dimethylglycine solution diluted with ammonium formate or an isotopically labeled choline solution diluted with ammonium formate;
(4) eluent: the mobile phase A is an ammonium formate solution containing formic acid, and the mobile phase B is acetonitrile.
9. The diagnostic kit for fetal congenital heart disease according to claim 8, wherein said acetonitrile solution is an acetonitrile solution of 80% and an acetonitrile solution of 100% by mass; in the internal standard solution, the concentration of isotope labeled dimethylglycine or isotope labeled choline is 200-500ng/ml, and the concentration of ammonium formate is 10 mmol/L; in the mobile phase A, the mass percentage of formic acid is 0.005-0.02%.
10. A system for detecting dimethylglycine in serum, which is characterized by comprising:
the standard curve drawing module is used for drawing a quantitative standard curve by taking the peak area ratio of the dimethylglycine to the isotope labeled internal standard as a vertical axis and the concentration of the dimethylglycine as a horizontal axis based on the fact that the dimethylglycine is used as a standard product and the isotope labeled dimethylnucleotide or isotope labeled choline is used as an internal standard;
a separation module for extracting and eluting dimethylglycine 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 dimethylglycine 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.
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| CN117110627B (en) * | 2023-10-18 | 2024-01-09 | 天津云检医学检验所有限公司 | Markers for neonatal dry blood spot congenital heart disease and subtype assessment thereof |
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