34 results about "MTRR" patented technology

The invention provides a method for typing three sites of key enzyme genes (MTHFR and MTRR) for folic acid metabolism by utilizing a fluorescence labeling probe and combining with a multiple PCR method. An amplimer and a fluorescent probe are designed according to MTHFR and MTRR genes; three sections of DNA sequences are detected in the amplification of a PCR amplifier; fluorescence signals which are released in amplification and dissolution processes in a reaction system are detected; a result is judged in the manner of (1) judging a typing result according to a Tm value shown by a hybrid peak of wild type and mutant type standard plasmids and (2) simultaneously typing according to the magnitude of a fluorescence value of an amplification curve. The primer and the probe adopted by the invention are high in specificity and sensitivity; the detection for three sites is simultaneously performed; the operation is simple and the result is easily judged; the typing result is more accurate and reliable in the manner of twice calibrating typing. The method provided by the invention can be applied to the aspects of guiding the pregnant woman to orally take and supplement folic acid, prompting the high risk in cerebral apoplexy, coronary heart disease and venous thrombus, assessing the metabolic activity of folic acid, and the like.

The invention discloses an MTHFR and MTRR gene polymorphism detection primer group and a kit. Mutation primers and probes aiming at three genetic loci have the advantages of being high in specificity and sensitivity. The prepared kit can detect MTHFR677, MTHFR1298 and MTRR66 gene polymorphism conditions, operation is simple, the experimental period is short, and the primer group and the kit are safe, free of toxicity, low in cost and suitable for clinical large-scale use and popularization.

The present invention relates to the fields of biotechnology and medicine, and provides specific sequence specific primers-polymerase chain reaction primer sequences and a kit for simultaneously detecting human MTHFR gene polymorphism and human MTRR gene polymorphism, wherein the kit contains the specific primers, specific probes, an internal standard system, Taq enzyme and UNG enzyme. According to the present invention, the primers and the kit have advantages of strong specificity, high sensitivity, simple and rapid operation, high-throughput, safety, objective result interpretation, and the like.

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.

Disclosed herein are novel assays, systems and kits for selecting a treatment regimen for a subject with depression by identifying at least one nucleic acid polymorphism, e.g., but not limited to, at the MTHFR, MTR, or MTRR locus, and / or determining expression levels of peripheral biomarkers (e.g., SAM, SAH, and 4-HNE) in a test sample from a human subject. These biomarkers can be used to determine the effectiveness of treating a depressed subject with a folate-containing compound (alone or as an adjunct to an antidepressant). Additionally, these biomarkers can be used to select an appropriate treatment regimen for subjects with treatment-resistant depression (e.g., resistant to at least one selective serotonin reuptake inhibitor). Methods and compositions for treating a subject with depression and / or determining or improving the effectiveness of an antidepressant drug taken by a subject are also provided herein.

The invention relates to an application of molecular beacon probes and a melting curve technology in detection of gene polymorphism, in particular to a method for detecting MTHFR (methylene tetrahydrofolate reductase) and MTRR (methionine synthase reductase) gene polymorphism simultaneously, and further relates to amplification primers of MTHFR and MTRR genes, the molecular beacon probes and a kitcontaining the amplification primers and the molecular beacon probes. The method is simple and fast to operate, good in specificity, high in sensitivity, accuracy and flux and low in detection cost and has broad application prospect. According to the method, PCR amplification and detection are performed synchronously, the overall detection process does not require uncovering operation, so that detection period is shortened and detection efficiency is improved while detection cost is saved, and besides, risk of false positive caused by PCR product pollution is reduced.

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 discloses a human MTHFR and MTRR gene detection kit and an application of the human MTHFR and MTRR gene detection kit. The kit comprises the following components: a MTHFR 677 / MTHFR 1298 / MTRR 66 triplet detection liquid and the following optional components of a nucleic acid amplification reaction liquid, an enzyme mixed solution, a positive control and a blank control. The disclosedhuman MTHFR and MTRR gene detection kit can detect genotypes of a 677 locus and a 1298 locus of a MTHFR gene and a lotus 66 of a MTRR gene at the same time in a same reaction tube. In addition, the kit further has the advantages of strong specificity, high sensitivity, convenient operation, strong repeatability, rapid and objective detection result and the like. The provided in vitro detection ofthe human MTHFR and MTRR genes provides an effective technical means.

The invention discloses a human MTHFR and MTRR gene polymorphism detection primer, probe, test kit and method and belongs to the technical field of in-vitro nucleic acid detection. The two most common single nucleotide polymorphisms 677C>T and 1298A>C of the MTHFR and the common mutation site 66A>G of the MTHFR are designed for qualitative detection, and a specific primer containing reference genes beta globin and a probe are designed. The test kit includes a detection primer and probe combination, a reference substance, a PCR reaction liquid and the like. Three independent multiplex PCR reactions are conducted for qualitative detection for the three gene polymorphism sites, namely, MTHFR677C>T, MTHFR1298A>C, and MTHFR 66A>G respectively. The detection primer, the probe, the test kit and the method are strong in specificity, high in sensitivity, simple, rapid and convenient for large-scale application and popularization.

The invention provides an MTHFR and MTRR gene polymorphism detection primer and probe combination, which includes an MTHFR C677T, MTHFR A1298C and MTRR A66G gene polymorphism detection primer and probe combination. The invention further provides a detection kit for the MTHFR and MTRR gene polymorphism. The kit is prepared from the following components of a nucleic acid releasing agent, 2*PCR reaction mixture (including a thermally activated Taq DNA polymerase and a UDG enzyme), three sets of specific primers, three sets of specific probes and an internal standard system. The kit only needs tobriefly treat a 2 [mu]l blood sample at the room temperature to produce high-quality DNA for repeated detection, the detection specificity is good, the coincidence rate with a gold standard sequencingmethod is 100%, the sensitivity is high, accurate typing of 0.2ng genomic DNA can be achieved, the detection process is short in consumed time, the process from sample processing to detection resultscan be completed within 1 hour.

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 discloses primers and probes for detecting genes related to metabolic capacity of folic acid and belongs to the technical field of biomedicine and also provides a kit containing the primers and probes. The primers and probes for detecting the genes related to metabolic capacity of folic acid have good specificity and can improve detection specificity of SNP sites; when the kit is used for detecting polymorphism of MTHFR and MTRR genes, the accuracy of detection result and sequencing results is up to 100%, homozygous wild type, homozygous mutant type and heterozygosis type of three sites of a sample can be detected by three tubes simultaneously, and the detection efficiency is high; meanwhile, compared with judged results in delta Ct value method, the judged results are easierto judge, can be accurately typed by typing software and affirmed and rechecked through fluorescence quantitative PCR curves and are more accurate and reliable.

The invention discloses a composition for detecting polymorphism of MTHFR and MTRR gene and an application of the composition. Single nucleotide polymorphism sites in MTHFR and MTRR genes are detected through multi-PCR reaction of FAM, HEX and ROX (modified) channels. In order to improve the detection simplicity and specificity, one-tube typing detection is achieved through technical means of probe typing, and the whole operation and reaction processes are simplified; locked nucleic acid modification and secondary structure modification are carried out on a probe on the basis, the Tm value of the probe is increased, the probe combining specificity is further improved and finally the lowest detection limit, accuracy and specificity of the whole kit are improved through adjusting the ratio of a primer pair, the dosage of Taq enzyme and the dosage of magnesium ions; and the minimum detection concentration of the kit can reach 1ng / muL and the accuracy and the specificity can reach 100%.

The invention discloses a specific primer group for testing polymorphic sites of human MTHFR and MTRR genes, and the primer group comprises a specific primer and a probe for the sites of MTHFR (C677T), MTHFR (A1298C) and MTRR (A66G) genes; and the invention also relates to a kit for testing the polymorphic sites of human MTHFR and MTRR genes, the kit comprises lyophilized powder (10 x SSP buffer,dNTPs, MgCl2, Taq enzyme and trehalose) of PCR reaction liquid containing the primer and the probe, and the kit is stored at 2-30 DEG C away from light. The invention also relates to a test method fortesting the polymorphic sites of human MTHFR and MTRR genes, and the method avoids a procedure of DNA extraction, and directly adopts a microamount of whole blood for amplification, which requires alittle blood and reduces the pain of patients; and the method saves time and effort, and greatly reduces the economic expenditure.

The invention relates to MTRR gene polymorphism detection primers and a detection system and reagent kit adopting the same. The primers comprise the forward primer used under the condition that an MTRR gene c.66 locus is A, the forward primer used under the condition that the MTRR gene c.66 locus is G, the universal reverse primer corresponding to the MTRR gene c.66 locus, the forward primer used for detecting a beta-actin gene, and the reverse primer used for detecting the beta-actin gene. The nucleotide sequence of the forward primer used under the condition that the MTRR gene c.66 locus is A is shown in SEQ ID No.1. The nucleotide sequence of the forward primer used under the condition that the MTRR gene c.66 locus is G is shown in SEQ ID No.2. The nucleotide sequence of the universal reverse primer corresponding to the MTRR gene c.66 locus is shown in SEQ ID No.3. The nucleotide sequence of the forward primer used for detecting the beta-actin gene is shown in SEQ ID No.4. The nucleotide sequence of the reverse primer used for detecting the beta-actin gene is shown in SEQ ID No.5. The reagent kit used for MTRR gene polymorphism detection is capable of quickly and conveniently carrying out detection, high in sensitivity and accuracy and low in cost.

The invention provides a primer for detecting the polymorphism of a human MTRR gene by virtue of a molecular beacon method. The primer comprises a primer pair and a probe, wherein the primer pair is used for amplifying polymorphic sites of rs1801394 (c.66 is more than G) and has the following nucleotide sequences, and the probe is used for detection. The invention further discloses a kit and method for detecting the polymorphism of the MTRR gene by virtue of the primer and the probe and application of the primer. By utilizing the primer, the polymorphism of the MTRR gene rs1801394 (c.66 is more than G) can be rapidly detected, and the result is accurate and easy to interpret. Compared with a sequencing method, the method has the advantages that a PCR product does not need to subjected to aseries of complex subsequent treatments, PCR amplification and detection are synchronously carried out, and the uncovering operation is not required in the whole detection process, so that the risk that the PCR product is polluted is reduced, the detection time is greatly shortened, and the detection cost is lowered.

The invention relates to a method for detecting gene polymorphism by using a labeled probe and a specific primer sequence and a kit comprising a specific probe and a specific primer sequence and adopted in the method, which are suitable for the fields of biotechnologies and medicine. The kit provides the specific primer sequence and the labeled probe which are for simultaneously detecting polymorphisms of C677T loci and A1298C loci of human MTHFR genes and A66G loci of MTRR genes, wherein the kit contains Taq enzyme, the specific primer, the specific probe and an internal standard system. Theprimer and the kit are used for simultaneously detecting the polymorphisms of the C677T loci and A1298C loci of the MTHFR genes and the A66G loci of the MTRR genes, and have the advantages of strong specificity, high sensitivity, simple and rapid operation, high flux, safety, objective result interpretation and the like.

The invention discloses a human MTHFR and MTRR gene polymorphism detection primer, probe, test kit and method and belongs to the technical field of in-vitro nucleic acid detection. The two most common single nucleotide polymorphisms 677C>T and 1298A>C of the MTHFR and the common mutation site 66A>G of the MTHFR are designed for qualitative detection, and a specific primer containing reference genes beta globin and a probe are designed. The test kit includes a detection primer and probe combination, a reference substance, a PCR reaction liquid and the like. Three independent multiplex PCR reactions are conducted for qualitative detection for the three gene polymorphism sites, namely, MTHFR677C>T, MTHFR1298A>C, and MTHFR 66A>G respectively. The detection primer, the probe, the test kit and the method are strong in specificity, high in sensitivity, simple, rapid and convenient for large-scale application and popularization.

The invention discloses a kit for rapid and simultaneous detection of MTHFR and MTRR gene polymorphisms and a method thereof, and belongs to the technical field of gene detection. According to the kitfor rapid and simultaneous detection of MTHFR and MTRR gene polymorphisms, the gene polymorphisms include the following types of gene polymorphisms: MTHFR 677C being greater than T, MTHFR 1298A beinggreater than C and MTRR 66A being greater than G. The kit includes a sample treatment reagent used for rapid treatment of human whole blood samples, gene detection reagents which are premixed and single-packaged, a positive control and a negative control. The gene detection reagents comprise MTHFR and MTRR wild-type and mutant-type gene detection reagents. On the basis of guaranteeing the detection specificity and sensitivity, convenient, fast and efficient detection is realized, detection operating steps are reduced, detection reaction time is shortened, and the production cost and the detection cost are also reduced. The invention is beneificla to clinical detection, analysis and application.

The invention provides a kit used for detecting human MTHFR and MTRR gene polymorphism, and a detection method thereof. The kit is used for avoiding defects and insufficient in conventional gene polymorphism detection technology. A primer and a probe used for detecting MTHFR and MTRR gene polymorphism are provided; the primer and the probe can be used for detecting MTHFR gene 677C>T and 1298A>C site and MTRR gene 66A>G site polymorphism, and are high in sensitivity and specificity. The kit can be used for rapid, high efficiency, and accurate detection of human MTHFR gene 677C>T and 1298A>C site and MTRR gene 66A>G site polymorphism. The invention also provides the detection method used for detecting human MTHFR and MTRR gene polymorphism using the kit. The detection method is convenient inoperation, high in sensitivity and specificity, wide in suitable machine type range, real and reliable in detection results, and is convenient for popularization.

The invention provides a method for detecting polymorphism of folate metabolism-related genes. The method comprises the steps that specific amplification primers and single base extension primers of the C677T site and A1298C site of an MTHFR gene and the A66G site of an MTRR gene are synthesized in advance; to-be-detected sample genomic DNA is extracted from a to-be-detected sample to serve as a DNA template; a polymerase chain reaction PCR amplification system containing the specific amplification primers and the DNA template is prepared; a PCR apparatus is used for amplifying the PCR amplification system; digestion treatment is performed in sequence; a single base extension reaction system containing the single base extension primers is added into a reaction system for extension reactions in sequence, desalted purification treatment is performed in sequence, and a purified product is obtained; a detection instrument containing a mass spectrometer is used for detecting the purified product to determine the gene polymorphism of the C677T site and A1298C site of the MTHFR gene and the A66G site of the MTRR gene of the to-be-detected sample. The method for detecting the polymorphismof the folate metabolism-related genes can improve the detection efficiency of folate metabolism capacity in the body of a patient.

The invention discloses a method for carrying out typing detection of an A66G polymorphic site of an MTRR gene. An MTRR gene segment is amplified by adopting MTRR gene-specific primers SEQ1 and SEQ2; and meanwhile, MTRR gene-specific TAQman probes SEQ3-FAM-A and SEQ 4-VIC-G are designed in an amplification area defined by an MTRR gene-specific primer. The method is used for judging gene SNP site typing on the basis of gene-specific PCR combined with the TAQman probes; and by adopting MTRR-specific gene amplification and the TAQman probes, target gene typing detection is carried out on the basis of a quantitative PCR method of the TAQman probes, so that the method has the characteristics of being simple, fast, high in repeatability, high in sensitivity and good in specificity.

The invention discloses a kit of MTRR gene polymorphism rapid test. The kit comprises PCR reaction liquid; the PCR reaction liquid is prepared from 0.04-0.09 U / microliter of DNA polymerase, 0.1-0.5 mM of dNTPS, 0.5-1.5 X of 5X PCR buffer solution, 1-2.5 mM of MgCl2, 0.0005-0.015%(w / v) of lauryl sodium sulfate, 0.001-0.03%(w / v) of polyethylene glycol octylphenol ether, 0.2-1.0 muM of upstream primer, 0.2-1.0 muM of downstream primer, 0.3-0.5 muM of mutant molecular beacon and 0.3-0.7 muM of wild molecular beacon, and is used for testing cell samples. The technical problem to be solved is to provide a primer, molecular beacon and kit of MTRR gene polymorphism rapid test and a testing method thereof through which sampling can be conducted in the painless environment and which are simple in operation, low in cost, rapid in test, short in time and high in success rate, and accordingly guidance is provided for individualization folic acid use, and the fetal anomaly risk is reduced.

The invention discloses a one-step RT-qPCR human MTHFR and MTRR gene polymorphism expression detecting kit. The one-step RT-qPCR human MTHFR and MTRR gene polymorphism expression detecting kit comprises an MTHFR-C677T reaction buffer, an MTHFR-A1298C reaction buffer, an MTRR-A66G reaction buffer, a positive control, a negative control and a One step enzyme mixed liquor. The one-step RT-qPCR human MTHFR and MTRR gene polymorphism expression detecting kit is based on drug metabolism genetics as well as the advantages of high sensitivity and high specificity of fluorescent PCR probe technique and can directly detect translated products of expression areas for RNA (ribonucleic acid) sequences of MTHFR and MTRR, thereby, compared with existing DNA-real-time (deoxyribonucleic acid-real-time) fluorescent PCR methods, being closer to detection of whether expression proteins, namely, MTHFR and MTRR exist, and being higher in specificity of detecting results.

The invention discloses an MTRR (5-methyltetrahydrofolate-homocysteine methyltransferase reductase) gene mutation detection liquid chip and specific primers. The liquid chip mainly comprises each ASPE (allele specific primer extension) primer formed by tag sequences at 5' terminal and specific primer sequences aiming at target gene mutation sites at 3' terminal, microspheres coated by anti-tag sequences as well as amplification primers, wherein the specific primer sequences are SEQ ID NO.13 and SEQ ID NO.14 aiming at G28905A sites, SEQ ID NO.15 and SEQ ID NO.16 aiming at C1783T sites, SEQ ID NO.17 and SEQ ID NO.18 aiming at A66G sites, SEQ ID NO.19 and SEQ ID NO.20 aiming at A1049G sites, SEQ ID NO.21 and SEQ ID NO.22 aiming at C1243T sites and / or SEQ ID NO.23 and SEQ ID NO.24 aiming at C1349G sites. The liquid chip has the advantages that the coincidence rate of the detection results of the detection liquid chip provided by the invention and a sequencing method is as high as 100%; and parallel detection of wild types and mutant types of a plurality of mutation sites is achieved.

The invention discloses a kit for simultaneously detecting human MTHFR and MTGG gene sequences. The kit comprises specific amplification primers and probes for identifying C677T, A1298C, G1793A and T1317C sites of MTHFR genes and A66G site mutations of MTRR genes. Compared with other existing detection products, the kit can accurately judge the genotype of samples, and it is confirmed that the samples are heterozygous mutations or homozygous mutations.

The invention discloses a method for detecting folic acid metabolism related gene mutation through combination of overlap extension PCR and Sanger sequencing and primer combination for detecting folicacid metabolism related gene mutation. A nucleotide sequence of the primer combination is shown as SEQ ID NO.1 to 8, and the primer combination is utilized to establish the detection method disclosedby the invention; according to the method disclosed by the invention, the overlap extension PCR and the Sanger sequencing are combined to detect folic acid metabolism related gene locus single nucleotide polymorphism, only once PCR reaction and once sequencing reaction are needed in the whole process, PCR reaction times and sequencing reaction times are remarkably reduced, and 6 mutation sites related to folic acid metabolism of MTHFR and MTRR genes can be detected; the method can be further utilized to find new variation points in amplified fragments. As the detection technology is applied to detecting folic acid metabolism related gene locus single nucleotide polymorphism, important clinical significance is achieved; thus, not only are operation steps simplified and is detection cost reduced, but also the detection efficiency is improved; the method is worth of popularizing.