A system for analyzing MTHFR gene polymorphisms in glucose intolerance during pregnancy

Abstract

A system for the analysis of MTHFR gene polymorphisms (C677T & A1298C) in glucose intolerance during pregnancy, consisting of: a) a sampling unit configured for taking peripheral blood samples from pregnant women; b) a sample storage unit configured to store blood samples in EDTA tubes at 4 °C; c) a DNA extraction unit configured to isolate genomic DNA from blood samples using the Qiagen kit and to store isolated samples at -20 °C; d) a PCR amplification unit configured to amplify the MTHFR gene regions C677T and A1298C using specific primer sequences in a reaction volume of 15 µl; e) a restriction digestion unit configured to digest amplified PCR products with the restriction enzymes HInfi and Mbo II by RFLP for 16 hours at 37 °C; f) an electrophoresis analysis unit configured to analyze digested RFLP products by polyacrylamide gel electrophoresis (PAGE) to identify restriction sites in amplified DNA products; and g) a data analysis unit configured to calculate genotype and allele frequencies using the Hardy-Weinberg equilibrium and the chi-square test.

Description

AREA OF INVENTION

[0001] The present disclosure relates to a system for analyzing MTHFR gene polymorphisms in glucose intolerance during pregnancy. In particular, the present invention provides a system configured to identify MTHFR gene polymorphisms (C677T&A1298C) associated with gestational diabetes. BACKGROUND OF THE INVENTION

[0002] Gestational diabetes mellitus (GDM) is a glucose intolerance disorder that develops during pregnancy and affects approximately 15% of pregnant women worldwide. The prevalence is even higher in Asian populations, at 17–20%. The condition causes significant complications for both mother and fetus, including an increased risk of preeclampsia, cesarean section, macrosomia, and neonatal hypoglycemia.

[0003] The methylenetetrahydrofolate (MTHFR) reductase gene encodes an important enzyme in one-carbon metabolism (OCM) that regulates folate and vitamin B12 synthesis in women of childbearing age. Specific genetic polymorphisms in the MTHFR gene, particularly the variants C677T (rs1801133) and A1298C, reduce enzyme activity and increase homocysteine ​​levels, potentially influencing the development of gestational diabetes through disrupted folate metabolism pathways.

[0004] Current diagnostic approaches for assessing the risk of gestational diabetes lack comprehensive genetic screening options to identify susceptible populations based on MTHFR polymorphisms. Existing systems do not integrate the molecular analysis of these specific gene variants with clinical and hematological parameters to enable comprehensive risk stratification for gestational diabetes mellitus.

[0005] Therefore, there is a need for an integrated analysis system capable of detecting MTHFR C677T and A1298C gene polymorphisms in pregnant women while simultaneously correlating genetic data with clinical parameters to assess susceptibility to GDM and provide enhanced diagnostic options for pregnancy management. Summary of the invention

[0006] This disclosure relates to a system for the analysis of MTHFR gene polymorphisms in glucose intolerance during pregnancy. The system is configured for the analysis of MTHFR gene polymorphisms (C677T & A1298C) in pregnant women to assess glucose intolerance during pregnancy. The system comprises several functional units, including sample collection, DNA extraction, PCR amplification, restriction enzyme digestion, electrophoresis analysis, and comprehensive data analysis functions for determining genetic risk factors associated with the development of gestational diabetes mellitus.

[0007] The present disclosure aims to provide a system for the analysis of MTHFR gene polymorphisms in glucose intolerance during pregnancy. The system comprises: a sample collection unit for obtaining peripheral blood samples from pregnant women; a sample storage unit for storing blood samples in EDTA tubes at 4 °C; a DNA extraction unit for isolating genomic DNA from blood samples using a Qiagen kit and storing isolated samples at -20 °C; a PCR amplification unit for amplifying the MTHFR gene regions C677T and A1298C using specific primer sequences in a reaction volume of 15 µl; and a restriction digestion unit for digesting amplified PCR products with the restriction enzymes HInfi and Mbo II by RFLP for 16 hours at 37 °C.an electrophoresis analysis unit for analyzing digested RFLP products by polyacrylamide gel electrophoresis (PAGE) to identify restriction sites in amplified DNA products; and a data analysis unit configured to calculate genotype and allele frequencies using the Hardy-Weinberg equilibrium and the chi-square test.

[0008] One objective of the present disclosure is to provide a system for the analysis of MTHFR gene polymorphisms in glucose intolerance states during pregnancy.

[0009] Another objective of the present disclosure is to integrate molecular genetic analyses with the evaluation of clinical and hematological parameters in order to identify MTHFR gene polymorphisms associated with the risk of gestational diabetes mellitus in pregnant women.

[0010] Another objective of the present disclosure is the simultaneous detection of the genetic variants MTHFR C677T and A1298C using PCR-RFLP techniques in combination with statistical analysis tools to calculate odds ratios and risk assessments in cases of gestational diabetes mellitus.

[0011] Another objective of the present disclosure is to correlate data on genetic polymorphism with clinical parameters such as gestational age, platelet count and glucose tolerance level in order to improve the predictive capabilities for the development of gestational diabetes mellitus during pregnancy.

[0012] To further clarify the advantages and features of the present disclosure, the invention is explained in more detail with reference to specific embodiments illustrated in the accompanying drawing. This drawing merely shows typical embodiments of the invention and is therefore not to be understood as limiting its scope. The invention is described and explained more precisely and in greater detail with reference to the accompanying drawing. BRIEF DESCRIPTION OF THE FIGURE

[0013] These and other features, aspects, and advantages of the present disclosure will be better understood if the following detailed description is read with reference to the accompanying drawing, in which the same symbols consistently represent the same parts. The following applies: Fig. Figure 1 shows a block diagram of a system for analyzing MTHFR gene polymorphisms in glucose intolerance states during pregnancy according to an embodiment of the present disclosure.

[0014] Experts will also recognize that the elements in the drawing are shown for the sake of simplicity and are not necessarily to scale. For example, the flowcharts illustrate the process by highlighting the main steps to enhance understanding of the aspects of this disclosure. Furthermore, with regard to the design of the device, one or more components of the device may be represented in the drawing by conventional symbols, and the drawing may show only the specific details relevant to understanding the embodiments of this disclosure, so as not to clutter the drawing with details that are readily apparent to those skilled in the art after reading this description. DETAILED DESCRIPTION:

[0015] For a better understanding of the inventive principles, reference is made below to the embodiment shown in the drawing, which is described in specific terminology. However, this does not limit the scope of the invention. Changes and further modifications of the illustrated system, as well as further applications of the inventive principles, are possible, as would normally occur to a person skilled in the art in the field of invention.

[0016] It is clear to the person skilled in the art that the preceding general description and the following detailed description are exemplary and explanatory of the invention and are not intended as a limitation of it.

[0017] References in this specification to “an aspect”, “another aspect”, or similar expressions mean that a particular feature, structure, or property described in connection with the embodiment is included in at least one embodiment of the present disclosure. Therefore, occurrences of the expressions “in one embodiment”, “in another embodiment”, and similar expressions in this specification may all refer to the same embodiment, but need not.

[0018] The terms "includes," "include," or other variations thereof are intended to cover non-exclusive inclusion, such that a process or method that includes a list of steps may not only contain those steps but may also include other steps not expressly listed or inherent in such process or method. Likewise, the statement "includes..." in the case of one or more devices, subsystems, elements, structures, or components does not, without further limitations, preclude the existence of other devices, subsystems, elements, structures, components, or additional devices, subsystems, elements, structures, or components.

[0019] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as understood by a person skilled in the art in the field of the invention. The system, methods, and examples provided here serve only for illustration and are not to be construed as a limitation.

[0020] Embodiments of the present disclosure are described in detail below with reference to the attached drawing.

[0021] Fig. Figure 1 shows a block diagram of a system (100) for analyzing MTHFR gene polymorphisms in glucose intolerance states during pregnancy according to an embodiment of the present disclosure.

[0022] Referring to Fig.1 The system (100) comprises: a) a sampling unit (102) for collecting peripheral blood samples from pregnant women; b) a sample storage unit (104) for storing blood samples in EDTA tubes at 4 °C; c) a DNA extraction unit (106) for isolating genomic DNA from blood samples using a Qiagen kit and storing isolated samples at -20 °C; d) a PCR amplification unit (108) for amplifying the gene regions MTHFR C677T and A1298C using specific primer sequences in a reaction volume of 15 µl; e) a restriction digestion unit (110) for digesting amplified PCR products with the restriction enzymes HInfi and Mbo II by RFLP for 16 hours at 37 °C; f) an electrophoresis analysis unit (112) for analyzing digested RFLP products by polyacrylamide gel electrophoresis (PAGE) to identify restriction sites in amplified DNA products;and g) a data analysis unit (114) configured to calculate genotype and allele frequencies using the Hardy-Weinberg equilibrium and the chi-square test.;

[0023] In one embodiment, the sampling unit (102) is configured to take 3 ml of peripheral blood samples specifically from pregnant women with a gestational age of 20 weeks and an age between 18 and 45 years.

[0024] In one embodiment, the PCR amplification unit (108) is configured to perform PCR amplification in a total volume of 15 µl with 8 µl DH2O, 4 µl Taq mix, 1 µl forward primer, 1 µl reverse primer and 1 µl DNA.

[0025] In one embodiment, the PCR amplification unit (108) is configured to perform a PCR cycle with DNA denaturation for 5 minutes at 94 °C, annealing for 1 minute at 58.35 °C for MTHFR C677T and 50 °C for A1298C gene polymorphisms, extension for 1 minute at 72 °C and final extension for 5 minutes at 72 °C.

[0026] In one embodiment, the system (100) also includes a unit (116) for recording clinical data, which is configured to record primary data such as hemoglobin levels, fasting blood glucose, random blood glucose, and blood glucose levels 1 hour and 2 hours after eating.

[0027] In one embodiment, the system (100) also includes a unit (118) for analyzing hematological parameters, configured to analyze secondary data including red blood cells, white blood cells, hematocrit, platelet count, mean cell volume, mean cell hemoglobin, mean cell hemoglobin concentration, lymphocytes, polymorphonuclear cells, monocytes, eosinophils and basophils.

[0028] In one embodiment, the data analysis unit (114) is configured to perform a statistical analysis using Fisher's exact test and Student's independent t-test with a significance level of p<0.05.

[0029] In one embodiment, the system (100) further comprises a risk assessment unit (120) configured to calculate ratios for MTHFR A1298C and MTHFR C677T gene polymorphisms associated with the development of gestational diabetes mellitus.

[0030] In one embodiment, the system (100) also includes a correlation analysis unit (122) configured to determine associations between gestational age, platelet count and MTHFR gene polymorphisms in the risk assessment of gestational diabetes mellitus.

[0031] In one embodiment, the restriction digestion unit (110) is configured to use the polymerase chain reaction restriction fragment length polymorphism technique for the molecular analysis of MTHFR C677T and MTHFR A1298C gene polymorphisms.

[0032] The present invention relates to a system specifically designed to investigate the association between MTHFR gene polymorphisms and glucose intolerance during pregnancy. The system integrates several specialized functional units to provide comprehensive genetic and clinical analysis capabilities for risk assessment of gestational diabetes mellitus. The system includes a sample collection unit specifically configured for collecting peripheral blood samples from pregnant women within defined parameters, including gestational age and demographic specifications. The collected samples are stored under controlled conditions in a dedicated sample storage unit, which maintains sample integrity at optimal temperatures using EDTA tubes at 4°C.A central component of the system is the DNA extraction unit, which isolates genomic DNA from collected blood samples using the standardized Qiagen kit method. The extracted genetic material is then stored at -20 °C to ensure DNA stability for molecular analysis. This extraction function guarantees high-quality genetic material for subsequent amplification and analysis processes. The PCR amplification unit is a critical component of the system and is used to amplify specific MTHFR gene regions corresponding to the C677T and A1298C polymorphisms. The unit operates with precise reaction parameters and uses a 15 µl reaction volume with specific primer sequences and controlled thermal cycle conditions.The amplification process includes denaturation at 94 °C, annealing at variant-specific temperatures, and extension phases to ensure optimal amplification of the target gene sequences. Following amplification, the restriction digestion unit processes the PCR products using the restriction enzymes HInfi and Mbo II via restriction fragment length polymorphism techniques. The digestion process takes place under controlled incubation conditions at 37 °C for 16 hours and enables precise cleavage of the amplified DNA products at specific recognition sites characteristic of the target polymorphisms. The electrophoresis analysis unit provides visualization and analysis capabilities via polyacrylamide gel electrophoresis, allowing for the identification and characterization of the restriction fragments generated during the digestion process.This unit facilitates the accurate determination of genetic variants based on the fragment patterns observed after electrophoretic separation.

[0033] In addition to its molecular analysis capabilities, one embodiment of the system includes comprehensive data acquisition and analysis units. The clinical data acquisition unit records primary parameters such as hemoglobin levels, fasting blood glucose, blood glucose measurements, and postprandial blood glucose levels. A separate hematological parameter analysis unit processes secondary data such as complete blood count parameters, cell indices, and differential counts, which may correlate with the development of gestational diabetes mellitus. The data analysis unit features sophisticated statistical analysis functions, including Hardy-Weinberg equilibrium calculations, chi-square tests, Fisher's exact test, and independent Student's t-tests. These analytical functions enable a comprehensive evaluation of genotype and allele frequencies while simultaneously determining the statistical significance of observed associations.The advanced analysis components include a risk assessment unit that calculates odds ratios for specific MTHFR polymorphisms, enabling a quantitative evaluation of the risk of gestational diabetes mellitus associated with identified genetic variants. The correlation analysis unit further enhances the system's capabilities by identifying relationships between genetic factors, clinical parameters, and the development of gestational diabetes mellitus. Designed for large-scale studies, the system offers configuration options for analyzing extensive sample populations, including both cases of gestational diabetes mellitus and healthy control groups. This scalability ensures the statistical power required to detect significant associations and generate reliable risk assessment data.

[0034] In one implementation, the system is configured to receive clinical and molecular data from a defined population of pregnant women. A total of 304 pregnant women were selected for data collection, including 152 with gestational diabetes mellitus (GDM) and 152 healthy controls. The control group consisted of normal pregnant women without diabetes or other complications up to delivery. The system specifically considers selection parameters, such as a gestational age of 20 weeks, an age between 18 and 45 years, and a diagnosis of GDM according to the standards established by the International Association of Diabetes in Pregnancy Study Group (IADPSG). Exclusion criteria exclude participants with chronic conditions such as kidney disease, liver disease, or endocrine disorders affecting glucose metabolism.Pregnant women with pre-existing diabetes mellitus, multiple pregnancies, a history of gestational diabetes, lack of willingness to consent, communication barriers, severe comorbidities, non-pregnant individuals, preeclampsia, eclampsia, chronic hypertension, and hydatidiform mole were excluded from the study population.

[0035] In one implementation, the system's clinical data collection unit is configured to capture primary data, including hemoglobin (Hb) levels, fasting blood glucose (FBS), random blood glucose (RBG), 1-hour postprandial blood glucose (PPBS), and 2-hour PPBS values. The system also captures secondary data, including age, height, weight, systolic blood pressure, diastolic blood pressure, gestational age, red blood cell (RBC) count, white blood cell (WBC) count, hematocrit, platelet count, mean corpuscular volume (MCV), mean corpuscular hemoglobin (MCH), mean corpuscular hemoglobin concentration (MCHC), lymphocytes, polymorphonuclear cells, monocytes, eosinophils, and basophils. The sample collection unit is configured to receive 3 mL of peripheral blood samples from each participant. Samples are collected in EDTA tubes and stored at 4°C in the sample storage unit until processing.The DNA extraction unit is configured to isolate genomic DNA from blood samples using the Qiagen kit method and to store the isolated DNA samples at -20 °C for subsequent molecular analysis.

[0036] In one implementation, the PCR amplification unit is configured to amplify the gene regions MTHFR C677T and A1298C. Amplification is performed in a total reaction volume of 15 µl, comprising 8 µl DH₂O, 4 µl Taq mix, 1 µl forward primer, 1 µl reverse primer, and 1 µl DNA. The amplification process includes PCR cycle conditions consisting of an initial DNA denaturation step for 5 minutes at 94 °C, followed by annealing for 1 minute at 58.35 °C for MTHFR C677T and 50 °C for MTHFR A1298C, extension for 1 minute at 72 °C, and a final extension for 5 minutes at 72 °C. The restriction digestion unit is configured to digest the amplified PCR products with the restriction enzymes HInfi and Mbo II under restriction fragment length polymorphism (RFLP) conditions and incubate for 16 hours at 37 °C.The digested products are then transferred to the electrophoresis analysis unit, where the products are analyzed using polyacrylamide gel electrophoresis (PAGE) to identify restriction sites within the amplified DNA.

[0037] In this implementation, the system's data analysis unit is configured to statistically analyze the collected clinical and molecular data. Sample size was determined using the Open Epi web tool, with parameters set to a 99% confidence level, a margin of error of ±5%, and a sample size of 15%. This resulted in a calculated sample size of 304 individuals, evenly distributed between case (n = 152) and control groups (n = 152). Analytical procedures include the application of Fisher's exact test and Student's independent t-test to evaluate the primary and secondary data. Genotype and allele frequencies of the polymorphisms MTHFR C677T and MTHFR A1298C are determined using the Hardy-Weinberg equilibrium (HWE) with the chi-square test. Statistical significance is set at p < 0.05.

[0038] Analysis of primary and secondary clinical data revealed a significant association between age and GDM cases compared to controls (p = 0.000), while systolic and diastolic blood pressure showed no association. Weight (p = 0.120), hemoglobin (p = 0.663), erythrocyte count (p = 0.456), leukocyte count (p = 0.216), hematocrit (p = 0.362), MCV (p = 0.142), MCH (p = 0.209), lymphocytes (p = 0.922), monocytes (p = 0.318), and eosinophils (p = 0.314) did not differ significantly between cases and controls. Mean fasting blood glucose was higher in GDM cases; however, no significant association was observed (p = 0.4519). In gestational diabetes, the values ​​for gestational age, platelet count, polymorphism, blood glucose levels, blood glucose levels 1 hour after eating, blood glucose levels 2 hours after eating and fasting blood glucose were significantly higher.The chi-square p-values ​​showed statistical significance for gestational age (p = 0.000), platelet count (p = 0.000), polymorphism (p = 0.002) and glucose parameters (p = 0.000).

[0039] In MTHFR A1298C (rs180113), genotypes AA and AG were more frequent in cases (30.26% and 51.97%, respectively) than in controls (35.52% and 39.47%, respectively). The genotype chi-square p-value was not statistically significant (p = 0.078). Allele frequency analysis showed that the A allele was more frequent in controls (56.25%) than in cases (47.36%), while the G allele was increased in cases (52.63%) compared to controls (43.75%); this distribution was statistically significant (p = 0.034). The odds ratio for AG alleles suggested a low risk for GDM (OR = 0.7.95%, CI: 0.5021–0.9758). The analysis in dominant and recessive models showed no significant associations overall, neither in severe nor in mild forms, with OR values ​​for the dominant model of 0.648 (overall), 1.109 (severe) and 0.379 (mild) and OR values ​​for the recessive model of 0.787 (overall), 0.9319 (severe) and 0.710 (mild).

[0040] For MTHFR C677T (rs1801133), the CC genotype was more frequent in controls (59.86%) than in cases (44.73%), while CT was more frequent in cases (25.00%) than in controls (22.36%). The TT genotype was increased in cases (30.26%) compared to controls (17.76%). The genotype chi-square p-value was statistically significant (p = 0.014). The C allele frequency was higher in controls (71.05%) than in cases (57.23%), while the T allele was more frequent in cases (42.76%) than in controls (28.94%), with the allele distribution being statistically significant (p = 0.008). The odds ratio of the CT alleles suggested a low risk in GDM cases (OR = 0.5453, 95% CI: 0.3292, 0.9011). In the dominant model, MTHFR C677T showed strong associations with GDM overall (OR = 2.009), in severe cases (OR = 2.274), and in mild cases (OR = 1.701), with an overall chi-squared p-value of 0.007.In the recessive model, the association with GDM was lower but statistically significant (total chi-squared p = 0.005). The odds ratio for CT alleles indicated a high risk for GDM (OR = 3.285, 95% CI: 1.988–5.441), with statistical tests performed separately for severe and mild forms.

[0041] The results show that elevated platelet counts and polymorphism levels correlate significantly with an increased likelihood of developing gestational diabetes in pregnant women. Furthermore, age and gestational age prove to be important influencing factors for the development of gestational diabetes. In the context of gestational diabetes, an examination of lymphocytes, monocytes, eosinophils, and basophils is performed. The results indicate an association between the single nucleotide polymorphism (SNP) MTHFR C677T (rs1801133) and gestational diabetes. Moreover, the p-value is associated with the allele frequency in the MTHFR A1298C (rs180113) gene polymorphism, suggesting a slight association with the risk of gestational diabetes.

[0042] The drawing and the preceding description show examples of embodiments. Those skilled in the art will recognize that one or more of the described elements can be combined to form a single functional element. Alternatively, certain elements can be divided into several functional elements. Elements of one embodiment can be added to another embodiment. For example, the sequence of the processes described here can be changed and is not limited to the manner described here. Furthermore, the actions of a flowchart need not be implemented in the sequence shown; nor does it necessarily have to be performed by all actions. Actions that are not dependent on other actions can also be performed in parallel with the other actions. The scope of the embodiments is in no way limited by these specific examples.Numerous variations are possible, whether explicitly stated in the specification or not, such as differences in structure, dimensions, and material use. The range of embodiments is at least as broad as specified in the following claims.

[0043] Advantages, further benefits, and problem solutions have been described above with reference to specific embodiments. However, the advantages, benefits, problem solutions, and all components that can lead to an advantage, benefit, or solution occurring or becoming more apparent are not to be construed as critical, necessary, or essential features or components of individual or all claims. REFERENCES 100 A System for the Analysis of MTHFR Gene Polymorphisms in Glucose Intolerance During Pregnancy. 102 Sampling unit 104 Sample storage unit 106 DNA extraction unit 108 PCR amplification units 110 Restriction Digestive Unit 112 electrophoresis analysis unit 114 Data Analysis Unit 116 Unit for recording clinical data 118 Hematological Parameter Analysis Unit 120 risk assessment units 122 Correlation analysis unit

Claims

[1] A system for the analysis of MTHFR gene polymorphisms (C677T & A1298C) in glucose intolerance during pregnancy, consisting of: a) a sampling unit configured for taking peripheral blood samples from pregnant women; b) a sample storage unit configured to store blood samples in EDTA tubes at 4 °C; c) a DNA extraction unit configured to isolate genomic DNA from blood samples using the Qiagen kit and to store isolated samples at -20 °C; d) a PCR amplification unit configured to amplify the MTHFR gene regions C677T and A1298C using specific primer sequences in a reaction volume of 15 µl; e) a restriction digestion unit configured to digest amplified PCR products with the restriction enzymes HInfi and Mbo II by RFLP for 16 hours at 37 °C; f) an electrophoresis analysis unit configured to analyze digested RFLP products by polyacrylamide gel electrophoresis (PAGE) to identify restriction sites in amplified DNA products; and g) a data analysis unit configured to calculate genotype and allele frequencies using the Hardy-Weinberg equilibrium and the chi-square test. [2] System according to claim 1, wherein the sampling unit is configured to take 3 ml of peripheral blood samples specifically from pregnant women with a gestational age of 20 weeks and an age between 18 and 45 years. [3] System according to claim 1, wherein the PCR amplification unit is configured to perform PCR amplification in a total volume of 15 µl with 8 µl DH2O, 4 µl Taq mix, 1 µl forward primer, 1 µl reverse primer and 1 µl DNA. [4] System according to claim 1, wherein the PCR amplification unit is configured to perform a PCR cycle with DNA denaturation for 5 minutes at 94 °C, annealing for 1 minute at 58.35 °C for MTHFR C677T and 50 °C for A1298C gene polymorphisms, extension for 1 minute at 72 °C and final extension for 5 minutes at 72 °C. [5] System according to claim 1, further comprising a unit for recording clinical data configured to record primary data including hemoglobin levels, fasting blood glucose, random blood glucose and blood glucose levels 1 hour and 2 hours after eating. [6] System according to claim 1, further comprising a unit for analyzing hematological parameters configured to analyze secondary data including red blood cells, white blood cells, hematocrit, platelet count, mean cell volume, mean cell hemoglobin, mean cell hemoglobin concentration, lymphocytes, polymorphonuclear cells, monocytes, eosinophils and basophils. [7] System according to claim 1, wherein the data analysis unit is configured to perform a statistical analysis using Fisher's exact test and Student's independent t-test with a significance level of p<0.

05. [8] System according to claim 1, further comprising a risk assessment unit configured to calculate ratios for MTHFR A1298C and MTHFR C677T gene polymorphisms associated with the development of gestational diabetes mellitus. [9] System according to claim 1, further comprising a correlation analysis unit configured to determine associations between gestational age, platelet count and MTHFR gene polymorphisms in the risk assessment of gestational diabetes mellitus. [10] System according to claim 1, wherein the restriction digestion unit is configured to use the polymerase chain reaction restriction fragment length polymorphism technique for molecular analysis of MTHFR C677T and MTHFR A1298C gene polymorphisms.

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