124 results about "Methylenetetrahydrofolate reductase gene" patented technology

This invention features our discovery on usages of Methylenetetrahydrofolate Reductase (MTHFR) gene polymorphisms in predicting homocysteine (Hcy) level and / or incidence and prognosis of diseases associated with increased Hcy level in a subject, as well as predicting treatment effects of medicines in the category of Angiotension Converting Enzyme Inhibihor (ACEI) with and without combination with B Vitamins. This invention also features our discovery on laboratory and analytical methods that are essential to the above described usages of MTHFR gene polymorphisms. In addition, this invention features a kit that has translated the above discoveries into a practical and reliable tool that can be applied to accomplish the above described usages of MTHFR gene polymorphisms. This invention represents an important step in realizing personalized medicine, with the goal to tailor diagnosis, prevention and treatment strategy to meet individual needs.

The invention relates to a kit for simultaneously detecting statin metabolizing gene multisite mutations. The kit includes primer pairs and probe pairs for detecting APOE, SLCO1B1, CETP, ABCB1 and MTHFR gene sites. MGB (minor groove binder) probes and a real-time fluorescent PCR technique are applied in the kit. The kit has advantages of capability of being time saving and convenient, high sensitivity, capability of allowing positive and negative coincidence rates of a sample to be 99.5% or above, and the like. The kit is mainly used for personalized medication assisted diagnosis of statins such as simvastatin, atorvastatin and pravastatin.

The invention provides a kit for genotyping folate metabolism gene. The kit includes a PCR reaction liquid and a DNA hybrid membrane strip, the DNA hybrid film strip comprises a matrix vector and probes, four probes for MTHFR genes, two probes for MTRR genes and two probes designed for human genome are sequentially fixed on the matrix vector, each of the probes is an oligonucleotide sequence hybridized with the SNP locus of each of the corresponding gene, and the nucleotide sequences of the probes are represented by SEQ ID NO:1-8. The kit provided by the invention has the advantages of high sensitivity, rapid detection, good stability, realization of high or low flux detection, high flexibility, strong maneuverability for small clinical samples in some hospitals, less equipment investment and low cost.

The invention provides a primer, probe, fluorescent PCR kit and method for detecting nucleotide polymorphisms of two sites of c.665 and c.1286 in an MTHFR (Methylene Tetrahydrofolate Reductase) gene. The nucleotide types of the two polymorphic sites of c.665 and c.1286 in the MTHFR gene are detected on a real-time fluorescent quantitative PCR (Polymerase Chain Reaction) technology platform according to the principle of 'allelic specific PCR'.

The invention provides a primer pair, specific oligonucleotide fluorescence probe and kit for detecting the nucleotide polymorphism of a C677T locus of an MTHFR gene. The new specific primer pair and the corresponding fluorescence probe are designed, PCR amplification is performed by adopting an asymmetric PCR technology, melting curve analysis is performed after PCR amplification is finished, and a genetype is judged through the melting peak at specific temperature. According to the kit, the MTHFR gene can be amplified in a highly specific mode, detection of three kinds of genetypes of the C677T locus in a single-pipe PCR system can be completed, the detection specificity is high, the result is easy to judge and read, the operating steps are simple, the detection cost is low, the cycle is short, and the efficiency is high.

The invention discloses a polymorphism site genetype of MTHFR gene to predict individual cardiovascular disease and prognostic application in the medical domain, which detects polymorphism parting oligonucleotide of MTHFR gene and polymorphism site genetype through MTHFR gene to predict the individual cardiovascular disease and prognostic method and agent box.

The invention provides a 5-fluorouracil drug sensitivity gene chip detection kit, which can be used to parallelly and economically detect genes related to 5-fluorouracil drug sensitivity. The kit comprises an extraction solution, a hybridization buffer solution, a washing liquor, an amplifying solution, Taq enzyme and a gene chip, the amplifying solution contains a primer for amplifying four genes containing mutant sites, and the four genes include a DPD gene containing IVS14 A / G mutant sites, a GSTPi gene containing Ile105Val mutant sites, an MTHFR gene containing C677T mutant sites, and an XRCC1 gene containing Arg399Glu mutant sites. The 5 fluorouracil is a first-line anti-tumor chemotherapeutic drug, and the detection kit can be used for detecting the four gene mutation conditions sensitive to the 5 fluorouracil drug.

The invention discloses a kit for detecting whether mutation occurs at a relevant single nucleotide polymorphism (SNP) locus on a disease-causing gene of a neural tube defect of a neonatus. The kit mainly comprises a specific primer pair and a specific fluorescent probe pair which are used for detecting a No.rs1801133 SNP locus polymorphism genotype and a No.rs1801131 SNP locus polymorphism genotype on a methylenetetrahydrofolate reductase (MTHFR) gene and a No.rs1801394 SNP locus polymorphism genotype on a methionine synthase reductase (MTRR) gene, and the conventional fluorescent quantitative PCR reaction reagent. The kit is used for detecting a mutant of the disease-causing gene of the neural tube defect of the neonatus, can be used for quickly and conveniently searching a carrier with the disease-causing gene, can be applied to antenatal diagnosis and timely treatment, and reduces the morbidity of the neutral tube defect of the neonatus so as to fulfill the aims of promoting good prenatal and postnatal care.

The invention discloses a kit for investigating polymorphism of an MTHFR (Methylene Tetrahydrofolate Reductase) and / or MTRR (Methylenetetrahydrofolate Reductase) gene on the basis of a Taqman-MGB probe. The kit comprises a primer group and a probe, wherein the primer group is at least one selected from the following three groups: primers of SEQ ID NO.1 and SEQ ID NO.4 for an MTHFR gene at a C677Tsite, primers of SEQ ID NO.7 and SEQ ID NO.10 for an MTRR gene at an A1298C site, primers of SEQ ID NO.13 and SEQ ID NO.14 for an MTRR gene at an A66G site, primers of SEQ ID NO.22 and SEQ ID NO.19, primers of SEQ ID NO.29 and SEQ ID NO.15, and primers of SEQ ID NO.33 and SEQ ID NO.30. The kit disclosed by the invention has the advantages that three mutation sites can be detected simultaneously, high sensitivity is achieved, plasma as low as 10copies can be accurately detected, and oral cavity swabs which are too long in preservation time or relatively low in concentration can be still accurately detected.

The invention provides a biosynthesis method for increasing accumulation of L-5-methyltetrahydrofolate, and an L-5-methyltetrahydrofolate synthetase system co-expressed recombinant plasmid and a construction method and application thereof. The L-5-methyltetrahydrofolate synthetase system co-expressed recombinant plasmid comprises DHFR (dihydrofolate reductase) gene folA and a MTHFR (methylenetetrahydrofolate reductase) gene metF sequence. The biosynthesis method for increasing accumulation of L-5-methyltetrahydrofolate includes converting the L-5-methyltetrahydrofolate synthetase system co-expressed recombinant plasmid to accumulate an original strain of the L-5-methyltetrahydrofolate so as to obtain a recombinant strain, and fermenting the recombinant strain. The accumulation of the L-5-methyltetrahydrofolate in final fermentation product is evidently higher than that of the L-5-methyltetrahydrofolate in the original strain. Utilization rate of raw materials is increased, production cost and energy consumption are reduced, and a foundation for industrial biosynthesis of the L-5-methyltetrahydrofolate is laid.

The invention discloses application, a detection method and a kit of SNP of an METHFR gene guiding folic acid and related nutrient element intake. Because of providing different positive locuses of SNP gene types of folic acid metabolic capacity and related nutrient element intake, the reason generating impediment thereof is found, and the guidance for guiding individuals, especially pre and post natal women, to take folic acid and intake related nutrients is provided.

The invention discloses a kit, a PCR (polymerase chain reaction) primer and a nucleic acid hybridization membrane strip for folic acid metabolism genotyping. The nucleic acid hybridization membrane strip comprises a substrate and specific polymorphic detection probes fixed to the substrate, the polymorphic detection probes are in one-to-one correspondence to folic acid metabolism related gene polymorphic sites, and the related gene polymorphic sites include at least one of a c.677 site and a c.1298 site of an MTHFR (methylene tetrahydrofolate reductase) gene, a c.56 site and a c.66 site of an MTRR (methionine synthase reductase) gene, a -186 site, a c.2756 site and a +905 site of an MTR (methionine synthase) gene and a -551 site of a CBS (cystathionine-beta-synthetase) gene. Each polymorphic detection probe comprises 13-25 base sequences. The kit, the PCR primer and the nucleic acid hybridization membrane strip are easy for judgment of polymorphic heterozygotes, polymorphic homozygotes and the like and have advantages of high flux, high efficiency, low cost, convenience in use and the like.

The invention discloses a kit used for detecting susceptibility to femoral head necrosis. The kit detects six genes which have close relation with the femoral head necrosis, namely a vascular endothelial growth factor (VEGF) gene of vascular endothelial factors, a coagulation factor V (FV) gene of coagulation factors, a methylene tetrahydrofolate reductase (MTHFR) gene, a pamoxonase 1 (PON1) gene of lipid metabolism related gene, an apolipoprotein A1 (APOA1) gene and an apolipoprotein B (APOB) gene. The kit detects a group of genes and loci related to the susceptibility to the femoral head necrosis through specific primers and probes by combining a single nucleotide extension technology with a microarray chip technology, can clearly judge whether examinees carry femoral head necrosis susceptibility genes and screens a group susceptible to the femoral head necrosis from the examinees, so as to make the group change bad living habits and fulfill the aim of preventing the femoral head necrosis.

The invention discloses a kit for genetic detection of colorectal cancer. The kit comprises a specific primer pair and a specific probe pair which are used for detecting the mononucleotide polymorphism loci genotype of a 5,10-methylenetetrahydrofolate reductase gene (MTHFR), a glutathione-s-transferase T1 gene (GSTT1), a glutathione-s-transferase M1 gene (GSTM1), a cytochrome P450 2E1 gene (CYP2E1), and a mismatching repair enzyme gene (hMLH1), a fluorescent quantitative PCR general component, a PCR reaction component and the like. The kit can evaluate the genetic risk of individuals suffered from the colorectal cancer by detecting the polymorphism loci genotype of the genes closely related to the genetic risk of the colorectal cancer at the same time.

The invention discloses a specific primer for detecting fluorouracil medicament healing effect related gene mutation, a liquid phase chip thereof and a method thereof. The liquid phase chip comprises wild type primer pair and mutation specific ASPE primer pair which are respectively designed for each type of mutation position point, microballoon which respectively comprises specific anti-tag sequence, and a primer which is used for enlarging target sequence with DPYD, TS and MTHFR gene mutation position points. The liquid phase chip for detecting the fluorouracil medicament healing effect related gene mutation has extreme good signal-noise ratio, and cross reaction is basically unavailable between a designed probe and the anti-tag sequence. The designed ASPE primer has extreme good specificity and can exactly distinguish the each type of mutation position point. The detecting method has simple step, can detect six type mutation position points at one time, has convenient operation, and avoids various uncertain factors existed in many operation processes, thereby being capable of greatly heightening the detecting accuracy rate.

There is provided is a nucleotide primer set for LAMP amplification used for detecting genotypes of single-nucleotide polymorphisms C677T and A1298C of an MTHFR gene. There is also provided a nucleotide probe for detecting an amplification product amplified by the primer set according to the present invention. There is also provided a method of detecting the genotypes of the single-nucleotide polymorphisms C677T and A1298C in the MTHFR gene, by using the primer set according to the present invention.

The invention discloses a gene combination, a primer and a probe used for detecting the susceptibility of femoral head necrosis, and application. The gene combination comprises six genes, namely a VEGF gene of vascular endothelial growth factors, an FV gene and an MTHER gene of blood coagulation factors, a PON1 gene, an APOA1 gene and an APOB gene of lipid metabolism genes, which are closely related to the femoral head necrosis. The existence of a femoral head necrosis susceptibility gene of detected population is comprehensively detected and analyzed by detecting a group of genes and loci which are related to the susceptibility of the femoral head necrosis, utilizing a specific primer and the probe and combining single-nucleotide extending technology with microarray chip technology, and the femoral head necrosis susceptible population is screened from the population for changing improper living habits, so as to fulfill the aim of prevention.

There is provided is a nucleotide primer set for LAMP amplification used for detecting genotypes of single-nucleotide polymorphisms C677T and A1298C of an MTHFR gene. There is also provided a nucleotide probe for detecting an amplification product amplified by the primer set according to the present invention. There is also provided a method of detecting the genotypes of the single-nucleotide polymorphisms C677T and A1298C in the MTHFR gene, by using the primer set according to the present invention.

The present invention provides a primer set for specifically amplifying a target region in a MTHFR gene by a nucleic acid amplification method, a MTHFR gene amplification reagent containing the primer set, and use of the primer set.

The invention discloses an SNP detecting kit for methionine-folic acid metabolism key enzymes MTHFR and a use method thereof. Corresponding specific primers and fluorescence probes for detecting MTHFR C677T polymorphism and MTHFR A1298C polymorphism are added, a real-time fluorescence quantification PCR system is formed, and through a real-time PCR, technicists in the field determine and detect the genotype of a corresponding MTHFR gene locus through data collected by a fluorogenic quantitative PCR amplifier. The advantages of being fast and sensitive, facilitating judgment and the like are achieved, and the SNP detecting kit and the use method thereof are suitable for clinical detection and popularization.

The invention relates to a kit and a detection method for polymorphism detection of a methylenetetrahydrofolate reductase gene. The kit comprises a nucleic acid extraction reagent and a nucleic acid detection reagent, wherein the nucleic acid detection reagent comprises fluorescent probe-containing PCR reaction liquid, a negative control and a positive control; the nucleic acid extraction reagent is a DNA extract; the PCR reaction liquid comprises DNA polymerase, UNG enzyme, a primer, a probe and Mg<2+>; the probe is a molecular beacon probe; and the gene polymorphic site of MTHFR is a C677T polymorphic site. The detection method using the kit is easy and fast to operate, the use of consumables is reduced so as to reduce the detection cost, the detection result is accurate, and the occurrence of false positive result is reduced; by adding the negative control and positive control in the amplification step, the judgment of false positive and false negative can be effectively improved, and the detection precision is higher; since the hairpin structure of the molecular beacon probe is opened with certain force, the specificity to the mutant C677T polymorphic site at the measuring point is higher.

The invention provides a cardiovascular and cerebrovascular diseases susceptibility gene chip detection kit belonging to the gene chip technique field of clinical detection, which can be used to parallelly and economically detect thirteen genes related to the susceptibility of cardiovascular and cerebrovascular diseases. The kit comprises an extraction solution, a hybridization buffer solution, a washing liquor, an amplifying solution, Taq enzyme and a gene chip, the amplifying solution contains a primer for amplifying the thirteen genes containing mutant sites, and the thirteen genes respectively are: an Enos gene, an ADD1 gene, an APOB-2515 gene, an AT1R gene, an APOB-3500 gene, an APOB-4181 gene, an eNOS-894 gene, an FgB-148 gene, an APOE gene, a GNB3 gene, an AGT gene, an FVII gene and an MTHFR gene. A plurality of genes are related to the susceptibility of the cardiovascular and cerebrovascular diseases, and the detection kit can be used for detecting the related thirteen genes parallelly.

The invention relates to a detection method and a kit for coronary heart disease susceptibility loci. The method comprises the following steps: (1) extracting a genome deoxyribonucleic acid (DNA) of a sample, and amplifying a region containing rs1801133 loci in a methylene tetrahydrofolate reductase (MTHFR) gene exon of the sample, so as to obtain an amplification product; (2) detecting the genetype of single nucleotide polymorphism (SNP) loci rs1801133 in the amplification product by using a high resolution melting (HRM) analysis technology. The genetype analysis is carried out on the region containing the rs1801133 loci in the amplified MTHFR gene by adopting the HRM analysis technology. The method is simple and quick to operate, low in use cost and accurate in result, and can achieve batch inspection; whether the tested people carry with coronary heart disease susceptibility genes can be clearly judged by using the kit, the coronary heart disease susceptibility people can be screened out from people, the poor living habit is improved, and the target of prevention is achieved.

The invention provides a susceptible gene noninvasive detection kit for hypertension. The kit comprises a specific primer and a DNA sequencing primer of the pleomorphic locus gene type of three mononucleotides which comprise C825T(rs5443) on a GNB3 gene, C667T(rs1801133) on an MTHFR gene and T869C(rs1982073) on a TGF-B1 gene, a PCR reaction assembly, a PCR product purifying assembly and a DNA sequencing reaction assembly. The kit assesses a risk level of hypertension at the genetic level by detecting the pleomorphic locus gene type of three mononucleotides which are closely related to hypertension, and guides the population to prevent generation of the disease in advance. A sampling method comprises sampling by oral mucosa cells without pain and wound, and avoids cross infection. The gene noninvasive detection kit has advantages of accurate and reliable sequencing detection result, no need of expensive special Import Instrument, and easy popularization.

The invention discloses a method and a detection kit for preventing stroke by detecting folate dysbolism. Oral muomembranous cells of an examinee are collected, genomic DNA is extracted, the polymorphism of a 677C-T locus on a methylenetetrahydrofolate reductase gene (MTHFR) gene of the genomic gene is detected so as to determine whether folate dysbolism exists, and the examinee subjected to genic mutation is intervened through small dosage of folate and vitamin B in daily life, so that the stroke is prevented.

The invention provides a primer, a probe, a fluorescence PCR kit and a method for detecting nucleotide polymorphism at *6-type and *28-type loci of the UGT1A1 gene. According to the allele specific PCR principle, the nucleotide type at *6-type and *28-type polymorphism loci of the methylenetetrahydrofolate reductase UGT1A1 gene is detected on a real-time fluorescence quantitative PCR technical platform.

The invention discloses a folic acid metabolism gene polymorphism detecting primer, comprising (1) detection primers for polymorphic sites of MTHFR gene (rs1801133, C677T): an upstream primer as shownin SEQ ID No. 1, a downstream primer as shown in SEQ ID No. 2 and a probe as shown in SEQ ID No. 3; (2) detection primers for polymorphic sites of MTHFR gene (rs1801131, a1298C): upstream primers asshown in SEQ ID No. 4, downstream primers as shown in SEQ ID No. 5 and probes as shown in SEQ ID No. 6; (3) detection primers for polymorphic sites of MTRR gene (rs1801394, a66G): upstream primers asshown in SEQ ID No. 7, downstream primers as shown in SEQ ID No. 8 and probes as shown in SEQ ID No. 9. The invention also discloses the use of the detecting primer in preparing a folic acid metabolism gene polymorphism detecting reagent and a folic acid metabolism gene polymorphism detecting reagent kit. The detecting primer provided by the invention can simultaneously complete the genotyping ofthree loci in a single-tube PCR system, and has the advantages of simple and convenient operation, strong specificity, high sensitivity, high accuracy and easy interpretation.

The invention provides a reagent kit for noninvasive test of cervical cancer susceptibility gene, which comprises specific primers, DNA (deoxyribonucleic acid) sequencing primers, PCR(polymerase chain reaction) reaction components, PCR product purifying components, DNA sequencing reaction components and the like, wherein the specific primers and the DNA sequencing primers are used for testing four mononucleotide polymorphism locus genotypes of Arg72Pro(rs1042522) on gene P53, deletion of gene GSTM1 (null / present), deletion of gene GSTT1 gene (null / present) and C677T(rs1801133) on gene MTHFR. The reagent kit can be used evaluating the risk level of female cervical cancer proportion by testing the four mononucleotide polymorphism locus genotypes relative to generation of cervical cancer closely, so that women can be guided to prevent cervical cancer pointedly in terms of the gene according to the gene test result of each woman to be tested, and the proportion of cervical cancer can be reduced. The reagent kit can be used with the oral mucosa cell sampling which is painless and non-invasive, so that cross infection is avoided. In addition, the sequencing test results are accurate and reliable, purchasing of expensive import special instruments is omitted, and the reagent kit is easy to be applied and popularized.

The invention provides methods of predicting risk of developing a hyper-homocysteine-associated disease in a subject, based on genotyping of the methylenetetrahydrofolate reductase (MTHFR) gene, wherein the risk varies depending on whether the 677T polymorphism and the 1298C polymorphism are present in a cis configuration within a MTHFR gene or not. A preferred hyperhomocysteine-associated disease is myocardial infarction. Kits for predicting risk of developing a hyperhomocysteine-associated disease are also provided.

The invention provides a method for identifying gene polymorphism rs2274976 of human MTHFR. The method comprises the following steps: providing human genome DNA to be tested; using forward primer and reverse primer of the sequence near the gene polymorphism rs2274976 site of the amplified human MTHFR and uses the human genome DNA to be tested as template to perform PCR amplification reaction and obtain an amplification product; using restriction enzyme to perform enzyme cutting to the amplification product; and performing electrophoresis to the enzyme cutting product to identify the gene polymorphism of human MTHFR, wherein the penultimate basic group on the end of the forward primer 3' is a mismatching basic group C to form CCGG or ACCGGT structure with polymorphic G allele in the amplification product, or to form CCAGT structure with polymorphic A allele so that the corresponding incision enzyme can be used for enzyme cleavage. The method of the invention has simple operation, low cost and wide application range.