RU2742801C1 - Method for detecting diabetic fetopathy - Google Patents

Abstract

FIELD: medicine.

SUBSTANCE: method for detecting diabetic fetopathy of the fetus is proposed, including determining the content of at least one marker of diabetic fetopathy of the fetus, selected from the group consisting of alpha-1-microglobulin / bikunin precursor (AMBP), apolipoprotein A-II, apolipoprotein B-100, apolipoprotein C-III, complement factor H, prothrombin, kininogen-1, alpha-1-acid glycoprotein, plasminogen, inter-alpha-trypsin inhibitor H1 heavy chain and vitronectin, in the analyte, determination of the total protein content in the analyte, assessment of the proportion of at least one marker diabetic fetopathy of the fetus from the total protein content in the analyte.

EFFECT: invention improves accuracy of the analysis, as well as expanding the arsenal of existing methods of analysis in the diagnosis of diabetic fetopathy of the fetus.

5 cl, 5 dwg, 3 ex

Description

The present invention relates to the field of clinical diagnosis, and in particular to methods of analysis in the diagnosis of diabetic fetopathy.

According to the definition of the Russian Association of Obstetricians and Gynecologists, diabetic fetopathy is a neonatal disease that develops in newborns whose mothers suffer from diabetes mellitus during pregnancy, and is characterized by systemic damage, metabolic and endocrine dysfunctions. According to WHO data for 2017, depending on the region of residence and anthropometric data, from 10% to 25% of pregnant women are at risk of developing hyperglycemia during pregnancy. International Diabetes Federation (IDF) 2017 statistics [https://www.idf.org/our-activities/care-prevention/gdm] show that in the world up to 204 million women aged 20 to 79 live with various types of diabetes, while up to 21.3 million newborns have symptoms of hyperglycemia, which in 85.1% of cases was caused by gestational diabetes in the mother during pregnancy.

Despite the well-known etiology and pathogenesis of diabetic fetopathy, its diagnosis and prevention remain an acute problem at the present time. In particular, the imperfection of methods for accurate differential diagnosis of hyperglycemia does not allow timely treatment and prevent the development of diabetic fetopathy. The reason for the weak sensitivity of existing biochemical methods for diagnosing diabetic fetopathy is, first of all, the lack of reliable clinical markers that would allow diagnosing the development of fetopathy in the fetus.

A method of obtaining an analytical test system (MRM test) for multiplex identification and quantitative measurement of the content of proteins of interest in a biological sample by the content of the corresponding proteotypic marker peptides is known from the prior art, which includes the following stages:

1) identification of proteotypic marker peptide sequences unique for the protein;

2) selection of at least two marker proteotypic peptide sequences of the protein, the most suitable for research by monitoring multiple reactions;

3) prediction of fragments of peptides;

4) prediction of the MRM test in the form of a list of marker peptides, their fragments and the best detection parameters by monitoring multiple reactions;

5) synthesis of one or more marker peptides;

6) determination of the profile of transitions of one and several synthetic marker peptides by monitoring multiple reactions under conditions of an accelerated chromatographic gradient;

7) optimization of the MRM test in accordance with the obtained profiles, removal from the set of peptide fragments characterized by the lowest intensity values in the mass spectrum;

8) purification of the peptides synthesized in step 5);

9) preparation of a biological sample, providing for the removal of major proteins;

10) identification of the protein in a biological sample with the stabbing of synthetic peptides by monitoring multiple reactions under normal chromatographic gradient conditions based on the profiles obtained in step 6);

11) determination of the retention times of marker peptides with the introduction of the established values into MRM tests;

12) carrying out multiplex calibration measurements under normal chromatographic gradient conditions in a buffer solution of purified synthetic peptides against the background of a mixture of unrelated proteins and / or peptides;

13) quantitative measurement of the content of marker peptides in a biological sample under normal chromatographic gradient conditions with the introduction of synthetic peptides; and

14) judgment on the content of proteins of interest in a biological sample by the content of the corresponding proteotypic marker peptides, quantitatively determined at stage 13) [RF patent RU 2595835]. This method represents a general approach to the identification of proteome proteins and in itself is not applicable to the detection of diabetic fetopathy of the fetus.

The closest analogue of the claimed invention is a method for detecting diabetic fetopathy, which is a dynamic ultrasound examination of the fetus at various stages of pregnancy [Gestational diabetes: diagnosis, treatment, postpartum observation. Clinical guidelines (protocol) // Moscow, 2014]. However, often with the use of this method, the signs of diabetic fetopathy cannot be reliably detected, despite the recommendations to conduct this kind of research in the period starting from the 20th week.

In the second and third trimesters of pregnancy, with dynamic ultrasound (ultrasound), the so-called advanced fetal development syndrome can be registered, which is a consequence of the development of hyperinsulinemia. In this case, during an ultrasound study, the main indicator is an increase in the size of the fetus 2 weeks ahead of the real time, polyhydramnios, tissue edema and imbalance in the size of the fetus. Signs of diabetic fetopathy in an ultrasound study, in which special attention is paid to such a symptom as cardiopathy, head contour, swelling and thickening of the head, can be indicators of insulin therapy. When diagnosing fetopathy, the indicators of glucose in the mother's blood, the determination of insulin, as well as her predisposition to diabetes are also taken into account. For this reason, constant dynamic control of glucose levels throughout the gestation period is the main requirement for controlling the risk of developing fetopathy in accordance with the WHO recommendations from 2016. However, this method for detecting diabetic fetopathy does not give sufficiently reliable signs for diagnosis. In many cases, this method does not allow identifying signs of an emerging diabetic fetopathy, and hence, conducting effective corrective compensatory therapy. This leads to developmental complications in the postnatal period.

Proteomics is a field of molecular biology dedicated to the identification and quantitative analysis of proteins (in other words, high-throughput protein research). The totality of all cell proteins is called the proteome [James P. Protein identification in the post-genome era: the rapid rise of proteomics. (English) // Quarterly Reviews Of Biophysics. - 1997. - November (vol. 30, no. 4). - P. 279-331. PMID 9634650]. It is known from the prior art that the data obtained by proteomics methods can be used to form a deeper understanding of the causes of various diseases, for example, neurodegenerative, as well as to develop therapies. Proteomics is used to search for antigens suitable for creating new vaccines. Identification of proteins that are abnormally expressed in various cancers is of great importance for diagnostics using biomarkers, predicting and treating cancer [B. Nolting. The latest methods of studying biosystems. - M .: TEKHNOSPHERA, 2005 .-- S. 185 .-- 256 p. - ISBN 94836-044-X.].

In a clinical study of the proteome of pregnant women suffering from various types of diabetes mellitus (gestational diabetes mellitus, type 1 and 2 diabetes mellitus), markers were identified whose abnormal content in the proteome correlates with the development of fetal fetopathy. These markers make it possible to carry out differential diagnostics on a small volume of biological material (the volume of blood plasma used in the work was 2 μl) and to identify the primary causes of the development of diabetic fetopathy, which makes it possible to more accurately speak about its etiology and predict its course. The average values of the protein content in the proteome of women in the control group are presented in Table 1.

Table 2 shows the dynamic changes in the relative content of proteome proteins of the study participants in relation to the control group (pregnant women).

The data obtained made it possible to develop a method for minimally invasive monitoring of the development of pathology. Taking into account the correlation of various markers, the proposed method makes it possible to observe a full-scale proteomic picture of diabetic fetopathy. A relationship has been established between various marker proteins and lipid metabolism processes, signaling pathways of PPAR receptors, hemostasis processes, as well as proteins of the extracellular matrix and architecture of the extracellular space. The obtained data on interrelated cellular processes, taking into account semi-quantitative values, make it possible to determine the limits of the norm of the content of certain markers or their combinations in different study groups and to conduct differential levels of their content in groups with diabetic fetopathy.

Thus, the technical result of the claimed invention is to improve the accuracy of the analysis in the diagnosis of diabetic fetopathy of the fetus, as well as to expand the arsenal of existing methods of analysis in the diagnosis of diabetic fetopathy of the fetus.

The technical result of the invention provides a method of analysis in the diagnosis of fetopathy of the fetus, including:

- taking a blood sample from the mother;

- sample preparation of a mother's blood sample to obtain an analyte;

- determination of the content of at least one marker of diabetic fetopathy of the fetus in the analyte;

- determination of the total protein content in the analyte;

- assessment of the proportion of at least one marker of diabetic fetopathy of the fetus from the total protein content in the analyte.

Determination of the content of markers of diabetic fetopathy of the fetus in the analyte can be carried out by enzyme immunoassay, chromatography-mass spectrometry, or any other methods that allow determining the content of these markers in the analyte.

Determination of the total protein content in the analyte can be carried out by gas chromatography-mass spectrometry, photometry, or any other methods that allow determining the total protein content in the analyte.

The determination of the total protein content, as well as the content of markers of fetal diabetic fetopathy in the analyte can be carried out, for example, in the following way.

For the analysis, take 1-2 μl of a plasma sample of the mother's venous blood. To 2 μl (about 100 μg of protein) of the plasma sample add 10 μl of a denaturing solution consisting of 5 M urea, 15% acetonitrile, 0.5% deoxycholic acid sodium salt, 300 mM sodium phosphate buffer (pH 6.0) and 5 mM TCEP (tris- (2-carboxyethyl) phosphine). The solution is incubated at 45 ° C for 20 minutes to restore the sulfhydryl groups of cysteine amino acid residues. Then add 1.5 μl of a solution of 2% 4-vinylpyridine in 30% propan-2-ol to a final concentration of about 0.2%. The alkylation reaction with vinylpyridine is carried out at room temperature (22 ± 2 ° C) for 30 minutes. The volume of the sample is brought to 100 μl (the calculated protein concentration in the sample is 1 μg / μl) with 100 mM triethylammonium bicarbonate solution to achieve the reaction of the medium to pH = 7.5-8.5.

Next, enzymatic cleavage of proteins with trypsin, previously diluted in 30 mM acetic acid to a final concentration of 200 ng / μL, is carried out in two stages, at the first of which trypsin is added in a ratio of 1:50 (by weight of protein), and at the second stage - an aliquot of trypsin equivalent to a ratio of 1: 100 (by weight of protein). In both cases, the enzymatic hydrolysis reaction is carried out at a temperature of 38 ° C for 3 hours at each stage. The reaction is stopped by adding 2 μl of 10% formic acid solution.

The resulting solutions are dried under vacuum at a temperature of 30 ° C in the V-HV mode (vacuum, volatile compounds) for 40-60 minutes. The dry residue of the samples is reduced in 100 μl of a 0.5% formic acid solution (to a final concentration of peptides of 1 μg / μl). The sample obtained after reduction is subjected to gas chromatography-mass spectrometric analysis.

Chromatographic separation is performed on an Ultimate 3000 RSLC Nano system (Thermo Scientific) with a set injector loop volume of 5 μL and a sample washout factor of 3 (three full loop volumes before switching the valve to loading mode). The samples are constantly thermostated within the temperature range of + 5 ± 0.5 ° C. A 5 μL sample is applied to the column. The sampling rate was 12 μL / min, and the sample injection rate was 15 μL / min. Analytical pump flow rate 0.3 μL / min, pressure limited to 800 bar max (nominal 1000 bar max) with dynamic flow acceleration at a gradient of 0.996 μL / min ² . The viscosity coefficient on channel "A" is 101%, on channel "B" - 64.7%. Analytical time - 55 minutes, equilibration time 13 minutes, total analysis time 68 minutes. Separation of peptides is carried out in a gradient of the mobile phase "A" (aqueous solution of 0.01% formic acid, 0.03% acetic acid, pH = 2.5- 2.7 depending on the temperature of the solution within 18-20 ° C), and the mobile phase "B" (90% acetonitrile, 10% methanol, 0.01%) formic acid, 0.03% acetic acid) on the stationary phase Acclaim Pepmap ® (geometry 75 μm × 150 mm, 1.8 μm, 60A). The initial conditions of the elution gradient 98% "A": 2% "B", flow rate 0.3 μl / min, pressure on the column 310-320 bar, loading rate on the enrichment column - 15 μl / min. During chromatographic separation, a dynamic change in the flow rate up to 0.45 ml / min is used at a high relative content of the mobile phase "B" in order to increase the efficiency of washing the column from bound hydrophobic components.

Mass spectrometric analysis is performed on an Orbitrap Fusion high resolution hybrid mass spectrometer (Thermo Scientific) with an NSI ionization source in positive electrostatic ionization mode and dynamic potential at the entrance S-lens. Precursor ions are recorded using an orbital mass analyzer with a resolution of 60K in the range of m / z 425-1250 with a maximum ion accumulation rate of 15 milliseconds, or a minimum integration number of -4e5 ions. Selection by charge state z = 2 +… 6 +, active dynamic exclusion after six scans (n = 6) for 4 seconds lasting 180 seconds with +10 / -25 ppm asymmetric isolation. Peak-dependent scanning activation at the level of 40% of the chromatographic peak height with an average value of FWHM = 24 seconds and a signal-to-noise level of at least 150. Tandem scanning is carried out after isolation through a quadrupole with an isolation window of ± 0.75 Th with a shift of 0.5 Th. Activation type - HCD, relative activation energy - 27% with variable offset +/- 20%. The type of fragment ion detector is orbital with a resolution of 15K with a maximum accumulation time of 47 milliseconds, or the number of ion integration is 5e4.

The original data files (raw format obtained by recording data from mass spectrometers manufactured by Thermo Fisher) were converted to mgf format using the MSConvert program (ProteomeWizard). Identification is carried out against a database of amino acid sequences of proteins (database with amino acid sequences in FASTA format with restriction of the taxonomic group "Human", available on the open repository Uniprot, the version is updated every 3-4 months) with the inclusion of reversed sequences to verify the obtained identifications. For the search, trypsin was chosen as a hydrolyzing enzyme (specific cleavage at lysine and arginine residues, if there is no proline in position P1) with the maximum allowable number of missing internal cleavage sites not more than one.

The allowed charging states were from z = 2 + to z = 6 + with an acceptable accuracy of measuring the precursor ion of ± 5 ppm and an acceptable accuracy of measuring a fragment ion of ± 0.01 Da. The variable modification used for search and detection was Q / E deamidation, methionine single oxidation, and 4-hydroxyproline. The fixed modification was pyridylethylation with 4-vinylpyridine. The results are verified by the cut-off level FDR = 1% (false discovery rate) based on the total frequency of false positives for spectra corresponding to peptides (PSM, peptide-spectra match).

The qualitative composition of the proteome is determined by protein identifications, which were carried out using the Search Gui software version 3.3 (Compomics, Belgium) using the X! Tandem Vengeance search algorithm version 12.15.2. Quantitative analysis is carried out on the basis of the NSAF (normalized spectral intensity index) in a sample of proteins common for a group of samples after alignment with the intensity of the reference spectra. The sample of proteins of interest is normalized in terms of NSAF values by the sum of their partial contribution to the research subproteome. Dynamic changes in individual proteins are analyzed using the open Heatmapper software (Canada), and pair clustering is analyzed using the Pairwise Distance software (Canada). Changes are checked for correctness of clustering for a particular study group through Kendal's rank correlation coefficient. Dynamic changes are expressed numerically in FC units (folds change) or in logarithmic FC if the changes exceed FC> 10.

As an embodiment of the claimed invention, the protein Alpha-1-microglobulin / bikunin precursor (AMBP) can be used as a marker of fetal diabetic fetopathy in the analyte. In this case, the value of the relative content of the marker of diabetic fetopathy of the fetus is no more than 0.3% of the total protein content in the analyte.

Also, the protein Apolipoprotein A-II can be used in the analyte as a marker of diabetic fetopathy of the fetus in gestational diabetes mellitus. The value of the relative content of the marker of diabetic fetopathy of the fetus is at least 4.6% of the total protein content in the analyte.

Also, the protein Apolipoprotein B-100 can be used in the analyte as a marker of diabetic fetopathy of the fetus in gestational diabetes mellitus. In this case, the relative protein content in fetopathy is at least 0.14% of the total protein content in the analyte.

Also, as a marker of diabetic fetopathy of the fetus in gestational diabetes mellitus, the analyte can use, for example, the protein Apolipoprotein C-III. In this case, the relative content of the marker of diabetic fetopathy of the fetus is from 2.7 to 3.5% of the total protein content in the analyte.

Also, the complement factor H protein can be used in the analyte as a marker of diabetic fetopathy of the fetus in gestational diabetes mellitus. In this case, the relative content of the marker of diabetic fetopathy of the fetus is at least 0.25% of the total protein content in the analyte.

Also, the protein Prothrombin can be used in the analyte as a marker of diabetic fetopathy of the fetus in gestational diabetes mellitus of the mother. In this case, the relative content of the marker of diabetic fetopathy of the fetus is not less than 0.48% of the total protein content in the analyte.

Also, as a marker of diabetic fetopathy of the fetus in gestational diabetes mellitus of the mother in the analyte, for example, the protein Kininogen-1 can be used. In this case, the relative content of the marker of diabetic fetopathy of the fetus is from 0.43 to 0.47% of the total protein content in the analyte.

Also, as a marker of diabetic fetopathy of the fetus in gestational diabetes mellitus in the mother, the analyte can use the protein Alpha-1-acid glycoprotein 1. In this case, the relative content of the marker of diabetic fetopathy of the fetus is from 1.30 to 1.83% of the total protein content in the analyte ...

Also, Plasminogen protein can be used in the analyte as a marker of diabetic fetopathy of the fetus in gestational diabetes mellitus. In this case, the relative content of the marker of diabetic fetopathy of the fetus is from 0.35 to 0.42 %% of the total protein content in the analyte.

Also, as a marker of diabetic fetopathy of the fetus in gestational diabetes mellitus, the analyte can use the protein Inhibitor of inter-alpha-trypsin heavy chain H1. In this case, the relative content of the marker of diabetic fetopathy of the fetus is at least 0.63% of the total protein content in the analyte.

Vitronectin protein can also be used in the analyte as a marker of diabetic fetopathy of the fetus in gestational diabetes mellitus of the mother. In this case, the relative content of the marker of diabetic fetopathy of the fetus is no more than 0.48% of the total protein content in the analyte.

Alternatively, maternal venous plasma or serum can be used as a blood sample.

This method makes it possible to create complete and modular diagnostic panels for monitoring analysis and confirmatory analysis in the diagnosis of fetopathy.

In the case of early diagnosis and assessment of the risk of developing this pathology, it becomes possible to conduct timely corrective or compensatory therapy.

Carrying out a study in the format of a diagnostic panel according to the markers proposed within the framework of the invention or their combinations is possible, for example, in the format of an enzyme-linked immunosorbent assay (ELISA) or mass spectrometric (MS) analysis. At the same time, the advantages of MS analysis are indifference to the type of protein marker, higher sensitivity, high productivity (the ability to perform analysis in 15-30 minutes per patient), low cost of analysis, high accuracy in measuring the signal of markers, the possibility of compiling modular diagnostic panels without additional financial costs in a few minutes, depending on the purpose of the analysis (diagnosis, observation, confirmation) without compromising performance, as well as the possibility of quantitative analysis of the patient's proteomic profile aimed at assessing the risk of developing diabetic fetopathy.

The claimed method is illustrated by the following examples.

Example 1. Implementation of the claimed invention.

Patient M, 28 years old, the first normal pregnancy. Has been suffering from type 1 diabetes for 2 years. She is on a combination therapy (short + prolonged insulins). At the time of registration for pregnancy in the antenatal clinic, the disease is in the stage of subcompensation: the level of fasting blood glucose is 4.4 mmol / l, 1 hour after eating - 10.0 mmol / l. During pregnancy, the patient was under the supervision of an endocrinologist with glycemic control and correction of the therapy. The level of glycated hemoglobin (HbA1c) by trimester: 5.7% - 5.3% - 5.5%, respectively.

Ultrasound of the fetus in the 1st trimester - no features.

An ultrasound scan for a period of 26 weeks was normal.

For a period of 30 weeks, a proteomic study of blood plasma for the presence of fetopathy markers was carried out in accordance with the claimed method. The research results are shown in table 3:

The data obtained indicate a high risk of fetopathy in the fetus.

Ultrasound for a period of 32 weeks - a tendency to a large fetus, polyhydramnios, no signs of fetopathy were detected.

Ultrasound at 38 weeks - large fetus, estimated fetal weight 4100 ± 100 g, polyhydramnios, signs of fetopathy (hepatomegaly, double head contour).

Delivery at 38.5 weeks by caesarean section. A live, full-term girl was born, weight 4170 g, height 55 cm, Apgar score 8 \ 9, with signs of fetopathy (macrosomia, hepatomegaly) and morphofunctional immaturity.

Thus, the use of the claimed method improves the accuracy of the analysis in the diagnosis of diabetic fetopathy of the fetus.

Example 2.

Patient N, 24 years old, the first normal pregnancy. Has been suffering from type 1 diabetes for 8 years. She is constantly monitored by an endocrinologist, is on combination therapy (short + prolonged insulins). When registering for pregnancy in an antenatal clinic, the level of fasting glycemia is 5.0 mmol / l, 1 hour after eating - 18.0 mmol / l. During pregnancy, the patient was under the supervision of an endocrinologist with glycemic control and correction of the therapy. The level of glycated hemoglobin (HbA1c) in the 1st, 2nd and 3rd trimesters of pregnancy is 8.0% - 7.0% - 7.3%, respectively.

Ultrasound studies for a period of 12-13 weeks and 19-20 weeks - no pathology was revealed.

With an ultrasound examination for a period of 26 weeks - a tendency to a large fetus, moderate polyhydramnios, thickening of the cervical fold.

For a period of 28 weeks, a proteomic study of blood plasma for the presence of a fetopathy marker (Prothrombin) was carried out in accordance with the claimed method. The research results are shown in table 4:

The data obtained indicate a high risk of fetopathy in the fetus.

Ultrasound at 32 weeks - a tendency to a large fetus, macrosomia, polyhydramnios, thickening of the cervical fold, hepatomegaly.

Ultrasound at 36 weeks - large fetus, macrosomia, polyhydramnios, thickening of the cervical fold, double head contour, hepatomegaly, impaired uteroplacental blood flow.

Delivery at 38 weeks by caesarean section, a live boy was born, weight 4100 g, height 51 cm, Apgar score 8/9. The child has inhibition of reflexes, undescended testicles into the scrotum. The signs of fetopathy were revealed: macrosomia, hepatomegaly, tissue pastiness, moonlike face, short neck, short lower limbs, hypertrichosis, morphofunctional immaturity. An increased echogenicity of the periventricular zone was revealed during the NSS.

Thus, the use of the claimed method improves the accuracy of the analysis in the diagnosis of diabetic fetopathy of the fetus.

Example 3.

Patient K, 28 years old, second normal pregnancy. The first pregnancy ended in urgent spontaneous childbirth at 40 weeks, the child is healthy. At the initial visit to the antenatal clinic in the period of 10 weeks, the level of fasting glycemia was 4.9 mmol / L, further (OGTT) oral glucose tolerance test was not performed. At a period of 28 weeks, an increase in the level of fasting glucose up to 6.7 mmol / l was noted, the diagnosis of GDM was made, and diet therapy was prescribed. On the background of diet therapy, the level of fasting blood glucose is 5.3 mmol / l, 1 hour after eating - 7.0 mmol / l. For a period of 36 weeks, insulin therapy (long-acting insulins) was connected, the level of fasting glycemia was 5.2 mmol / L, 1 hour after a meal - 6.2 mmol / L.

During pregnancy, ultrasound studies were carried out at 21 weeks, 29 weeks, 36 weeks - no signs of fetopathy were revealed.

For a period of 29 weeks, a proteomic study of blood plasma for the presence of fetopathy markers was carried out in accordance with the claimed method. The research results are shown in table 5:

The levels of all tested proteins are within the normal range, which is an indicator of normal fetal development.

Delivery is urgent (at 40-41 weeks), spontaneous, through the natural birth canal. A live full-term girl was born, weight 3150 g, height 51 cm, Apgar scores 9 \ 9. There were no signs of fetopathy.

Thus, the use of the claimed method allows the analysis in the diagnosis of diabetic fetopathy of the fetus with high accuracy.

Appendix 2

An example of implementation of the claimed invention

Patient N, 31 years old, the first normal pregnancy. Has been suffering from type 1 diabetes for 3 years. She is on a combination therapy (short + prolonged insulins). At the time of registration for pregnancy, the level of fasting blood glucose was 4.7 mmol / l, 1 hour after eating - 9.7 mmol / l. During pregnancy, the patient was under the supervision of an endocrinologist with glycemic control and correction of the therapy. The level of glycated hemoglobin (HbA1c) by trimester: 5.4% - 5.2% - 5.4%, respectively.

Ultrasound of the fetus in the 1st trimester - no features.

An ultrasound scan for a period of 26 weeks was normal.

For a period of 26 weeks, a proteomic study of blood plasma for the presence of fetopathy markers was carried out in accordance with the claimed method.

The research results are shown in table 1:

The data obtained indicate a high risk of fetopathy in the fetus.

Ultrasound for a period of 30 weeks - a tendency to a large fetus, polyhydramnios, no signs of fetopathy were detected.

Ultrasound scan at 38 weeks - a large fetus, estimated fetal weight 4200 ± 100 g, polyhydramnios, signs of fetopathy (hepatomegaly, double head contour).

Delivery at 38 weeks by caesarean section. A live full-term boy was born, weight 4090 g, height 57 cm, Apgar score 8 \ 9, with signs of fetopathy (macrosomia, hepatomegaly) and morphofunctional immaturity.

Thus, the use of the claimed method improves the accuracy of the analysis in the diagnosis of diabetic fetopathy of the fetus.

Claims (10)

1. A method for detecting diabetic fetopathy of the fetus, including: - taking a blood sample from the mother; - sample preparation of a mother's blood sample to obtain an analyte; - determination of the content of at least one marker of diabetic fetopathy of the fetus selected from the group consisting of alpha-1-microglobulin / precursor bikunin (AMBP), apolipoprotein A-II, apolipoprotein B-100, apolipoprotein C-III, complement factor H, prothrombin , kininogen-1, alpha-1-acid glycoprotein, plasminogen, inhibitor of inter-alpha-trypsin H1 heavy chain and vitronectin, in analyte; - determination of the total protein content in the analyte; - assessment of the proportion of at least one marker of fetal diabetic fetopathy from the total protein content in the analyte, where, if: the value of the relative content of alpha-1-microglobulin / bikunin precursor (AMBP) is not more than 0.3% of the total protein content in analyte and / or the value of the relative content of apolipoprotein A-II is not less than 4.6% of the total protein content in the analyte and / or the value of the relative content of apolipoprotein B-100 is not less than 0.14% of the total protein content in the analyte and / or the value of the relative content of apolipoprotein C-III is from 2.7 to 3.5% of the total protein content in the analyte and / or the value of the relative content of the complement factor H is not less than 0.25% of the total protein content in the analyte and / or the value of the relative the content of prothrombin is at least 0.48% of the total protein content in the analyte and / or the value of the relative content of kininogen-1 is from 0.43 to 0.47% of the total about the protein content in the analyte and / or the value of the relative content of alpha-1-acid glycoprotein is from 1.30 to 1.83% of the total protein content in the analyte and / or the value of the relative content of plasminogen is from 0.35 to 0.42% of the total protein content in the analyte and / or the value of the relative content of inter-alpha-trypsin inhibitor H1 heavy chain is not less than 0.63% of the total protein content in the analyte and / or the value of the relative content of vitronectin is not more than 0.48% of the total the content of proteins in the analyte, determine the high risk of developing fetopathy in the fetus. 2. The method according to claim 1, characterized in that the determination of the content of markers of diabetic fetopathy of the fetus in the analyte is carried out by the method of enzyme immunoassay. 3. The method according to claim 1, characterized in that the determination of the content of markers of diabetic fetopathy of the fetus in the analyte is carried out by the method of chromatography-mass spectrometry. 4. The method according to claim 1, characterized in that the determination of the total protein content in the analyte is carried out by chromatography-mass spectrometry. 5. The method according to claim 1, characterized in that the determination of the total protein content in the analyte is carried out by the method of photometry.