54 results about "VKORC1 Gene" patented technology

The present invention relates to methods and compositions for predicting drug responses. In particular, the present invention provides methods and compositions for determining individualized Warfarin dosages based on the level of expression of the VKORC1 gene.

The invention discloses a primer and probe composition for detecting polymorphism of human CYP2C9 and VKORC1 genes, a kit and application. The primer and probe composition comprises three pairs of specific primers for amplifying CYP2C9*2, CYP2C9*3 and VKORC1 sites, and three specific fluorescent probes. The primers and the probes, disclosed by the invention, have high sensitivity, strong specificity and strong anti-interference capability; a manner combining an asymmetric PCR (Polymerase Chain Reaction) amplification and fluorescent probe melting curve analysis technologies is used for detecting the gene polymorphism; different gene types can be effectively distinguished according to the quantity and Tm value of a melting peak; a result is convenient, clear and objective to judge and read.Single-tube sampling can be used for simultaneously detecting 6 mutation types of 3 gene sites; the primer and probe composition is simple and convenient to operate and the detection efficiency is improved; a large batch of samples can be detected and clinical operation is facilitated.

The invention discloses a test kit for identifying single nucleotide polymorphism of target genes including a CYP2C9 gene and a VKORC1 gene in a biological sample. The kit comprises a first group of probes and a second group of probes, wherein a first probe in the first group of probes has a nucleotide sequence of which the similarity with any one of nucleotide sequences SEQ ID NO:3, 4 and 5 and a complementary sequence thereof is at least 80 percent; a second probe in the first group of probes has a nucleotide sequence of which the similarity with any one of nucleotide sequences SEQ ID NO:6, 7 and 8 and a complementary sequence thereof is at least 80 percent; a first probe in the second group of probes has a nucleotide sequence of which the similarity with any one of nucleotide sequences SEQ ID NO:11, 12 and 13 and a complementary sequence thereof is at least 80 percent; and a second probe in the second group of probes has a nucleotide sequence of which the similarity with any one of nucleotide sequences SEQ ID NO:14, 15 and 16 and a complementary sequence thereof is at least 80 percent. The reagent kit can also comprise a first primer pair and a second primer pair.

The invention discloses a CYP2C19, CYP2C9 and VKORC1 genotyping multiplex amplification system and detection kit. The amplification system comprises two forward primers and a reverse fluorescent primer of six SNPs (single nucleotide polymorphisms) respectively and can simultaneously amplify the six SNPs. The system is characterized by achieving one-tube amplification of three SNPs on the gene CYP2C19, two SNPs on the gene CYP2C9 and one SNP on the gene VKORC1 through multiple PCR (polymerase chain reaction). In particular, the amplification system can achieve direct amplification of blood and blood spot samples and dispenses with the step of extracting DNA (deoxyribonucleic acid). The amplification system can integrate the UDG-dUTP antipollution measure and can effectively prevent product pollution. The detection system is comprehensive in site detection, is simple and convenient to operate, has high specificity, high sensitivity and strong reliability, is low in cost and has the capacity of mass detection.

The present invention provides a human CYP2C9 and VKORC1 genetic polymorphism detection kit. The kit includes a PCR buffer, dNTP, MgCl2, 2 groups of specific primers, 2 groups of specific probes, an internal standard system, HotStart Taq enzyme and UNG enzyme. The detection kit for detecting CYP2C9 and VKORC1 genetic polymorphism has the advantages of high specificity, high sensitivity, easy and fast operation, high-throughput, security, and objective interpretation of the results.

The invention discloses a primer and a kit for detecting CYP2C9 and VKORC1 gene polymorphism and a PCR method of the primer and the kit. A detection CYP2C9 gene comprises a wild type specificity upstream primer, a mutant type specificity upstream primer and a first sharing downstream primer; the wild type specificity upstream primer has a SEQ No.17 sequence, the mutant type specificity upstream primer has a SEQ No.14 sequence, and the first sharing downstream primer has a SEQ No.16 sequence; a detection VKORC1 gene comprises a wild type specificity upstream primer, a mutant type specificity upstream primer and a second sharing downstream primer; and the wild type specificity upstream primer has a SEQ No.38 sequence, the mutant type specificity upstream primer has a SEQ No.35 sequence, and the second sharing downstream primer has a SEQ No.37 sequence. The kit has the beneficial effects of being simple, rapid and accurate in detection, low in cost and the like.

The invention discloses a kit for inspecting the curative effect of an individual Warfarin anticoagulant medicament. The kit comprises a specific primer pair, a specific fluorescent probe pair, a conventional fluorescence quantitative PCR assembly and the like, wherein the specific primer pair and the specific fluorescent probe pair are used for inspecting polymorphism sites of a CYP2C9 gene and a VKORC1 gene of targeted sites of the Warfarin medicament. The kit evaluates the curative effect of the individual Warfarin anticoagulant medicament by inspecting the polymorphism sites of the CYP2C9 gene and the VKORC1 gene of the targeted sites of the Warfarin medicament.

The invention discloses a primer, a probe and a kit for detecting gene polymorphism of CYP2C9 and VKORC1. The primer comprises a gene detection specificity primer sequence of three loci YP2C9*2, CYP2C9*3 and VKORC1 and a probe sequence (SEQ ID NO.1-12). An ARMS primer is used to differentiate field and mutant genes, so that the primer has the advantages of high sensitivity and high specificity and is suitable for various sample types and high in detection result accuracy. In addition, the ARMS primer is quick and simple to synthesize, low in synthesis cost and better in amplification effect, and production cost can be lowered effectively. The kit is high in detection speed and quite convenient, and the whole detection process only takes 90 minutes. The whole kit does not contain toxic and harmful substances, thereby being harmless to operating personnel and environment and suitable for wide application in clinic detection and the like.

The invention discloses a composition for detecting CYP2C19 and VKORC1 gene polymorphism and its application. The single nucleotide polymorphism site in the CYP2C19 and VKORC1 gene is detected through FAM, HEX, ROX (modified) three-channel multiple PCR reaction. In order to improve the detection simplicity and specificity, one-tube parting detection is realized through the technical manner of a probe parting, thus the whole operation and the reaction process are simplified; on this basis, locked nucleic acid modification and second-grade structure modification are performed on the probe to increase the Tm value of the probe, and further improve the probe combining specificity; finally, the lowest detection limit, accuracy and the specificity of the whole kit are improved through adjusting the mixture ratio of the primer pair, dosage of Taq enzyme, dosage of magnesium ion and others; the lowest detection concentration of the kit can reach 1 mg/mu l; the accuracy and the specificity can reach 100%.

The invention discloses a kit for detecting human CYP2C9 (Cytochrome P450 2C9) and/or VKORC1 (Vitamin K epoxide reductase complex subunit 1) gene polymorphism. The kit comprises a fluorescence detection probe I represented as SEQ ID NO:3, a fluorescence detection probe II represented as SEQ ID NO:6 and/or a fluorescence detection probe III represented as SEQ ID NO:9, wherein the fluorescence detection probe I comprises an amplimer for C43oT site of a CYP2C9 gene as well as a base; the fluorescence detection probe II comprises an amplimer for A1075C site of the CYP2C9 gene as well as a base; and the fluorescence detection probe III comprises an amplimer for G-1639A site of a VKORC1 gene as well as a base. Specific primers, fluorescence detection probes and an optimized detection system are adopted, the three gene types can be detected separately or synchronously, the detection linearity is wide, a 25 mu L detection system can detect 2ng-600ng human genomic DNA (deoxyribonucleic acid), the accuracy is high, and the detection result is completely consistent with that of the Sanger sequencing method.

The invention discloses a kit for detecting gene polymorphism of CYP2C9 and/or VKORC1. The kit comprises the following components: hot start taq DNA polymerase, a blood direct PCR buffer (10X), a primer pair, a probe pair, and a UNG enzyme. The kit is simple, high efficient, and safe: the product can be used for directly carrying out fluorescence PCR amplification of blood samples without nucleicacid extraction, purifying and other steps, risks of cross contamination of templates are reduced, and usage of phenol and other harmful reagents are avoided; the kit has high sensitivity: blood samples which are low to 2[mu]l can be accurately detected, and required sample amount is low; the specificity is good: specific primers and parting probes are designed aiming at CYP2C9C430T, CYP2C9A1075Cand VKORC1G-1639A, and corresponding polymorphism can be identified by specific amplification.

The invention relates to a primer pair and kit for detecting VKORC1 (vitamin K epoxide reductase complex subunit 1) genotyping by pyrosequencing, belonging to the technical field of in vitro nucleic acid detection. The primer pair comprises VKORC1 G1639A and VKORC1 C1173T forward amplification primers, VKORC1 G1639A and VKORC1 C1173T reverse amplification primers and VKORC1 G1639A and VKORC1 C1173T sequencing primers, wherein 5' terminals of the VKORC1 G1639A forward amplification primer and the VKORC1 C1173T reverse amplification primer are respectively subjected to biotin labelling. The kit comprises the amplification primers, a PCR (polymerase chain reaction) liquid 1, a PCR liquid 2, the sequencing primers, uracil DNA (deoxyribonucleic acid)glycosylase and Taq polymerase. The kit provided by the invention has the advantages of accurate detection results, high specificity, short detection period, simplicity in operation, capability of effectively meeting the requirements of clinical examination, capability of monitoring the reaction process in real time, short reaction time, sequencing of PCR products on a pyrosequencing instrument after the PCR products are simply treated, high throughput sample detection and higher sensitivity than gold standard methods, namely capillary electrophoresis sequencing methods.

Primer sets for amplifying two genes (the CYP2C9 gene and the VKORC1 gene) by a gene amplification method are provided, wherein the primer sets can amplify respective target regions of the two genes specifically and efficiently in the same reaction system simultaneously. Two pairs of primer sets are used including forward primers consisting of the base sequences of SEQ ID NOs: 5 and 29 as well as reverse primers consisting of the base sequences of SEQ ID NOs: 18 and 38, respectively. The use of these primer sets makes it possible to specifically amplify target regions including sites where polymorphisms to be detected are generated in the CYP2C9 gene and the VKORC1 gene, in the same reaction solution simultaneously.

The invention discloses a kit for rapid detection of polymorphism of a Warfarin metabolic enzyme gene by virtue of a pyrosequencing method. The kit consists of three pairs of specific amplification primers and three sequencing primers; the three pairs of specific amplification primers include two genes, namely a CYP2CP gene and a VOKRC1 gene, wherein the CYP2CP gene contains two pairs of primers; the three sequencing primers specifically relate to three single nucleotide polymorphisms, namely CYP2C9*2, CYP2C9*3 and VKORC1 (-1693); and the nucleotide sequences are as shown in SEQ ID NO.1-9. The detection method disclosed by the invention is not only high in sensitivity, but also direct in result, and simpler, rapider and more accurate in interpretation; the kit is capable of achieving accurate, rapid and high-throughput detection of the Warfarin metabolic enzyme gene; and the kit is relatively high in popularization and application values in personalized medicine.

The invention provides a primer group, a kit and a method for detecting gene polymorphism. The primer group comprises a primer pair for detecting an APOE gene, a CYP4F2 gene, a GGCX gene, an NPC1L1 gene, an NQO1 gene and a VKORC1 gene. When the primer group is used for performing multiplex PCR detection, mutation of at least one gene can be detected at the same time through one-time PCR reaction,so that the gene detection flux is improved.

The invention discloses a CYP2C9 and VKORC1 gene typing detection reagent kit. The CYP2C9 and VKORC1 gene typing detection reagent kit comprises a gene chip and various primers. The gene chip is provided with 4 mutation sites of CYP2C9 and VKORC1 genes and nucleotide sequence probes in normal contrast and complementation with the mutation sites, wherein the probes have the sequences shown in SEQ ID Nos. 1-8 or sequences complementary to the sequences shown in SEQ ID Nos. 1-8; the gene chip is further provided with an DNA sequence marked with biotin points, wherein the DNA sequence is shown in SEQ ID No.9. The primers have the sequences shown in SEQ ID Nos. 10-17. The CYP2C9 and VKORC1 gene typing detection reagent kit provides a CYP2C9 and VKORC1 gene typing joint detection platform, can achieve synchronous joint detection, improve detection specificity, lower the cost and shorten detection time, and has great significance in carrying out CYP2C9 and VKORC1 gene detection and mass screening.

The invention provides a primer probe combination for quantitative PCR (polymerase chain reaction) detection on CYP2C9 and VKORC1 gene polymorphism. The primer probe combination comprises three groupsof primers and double probes shown in sequence tables in the description. The invention further provides a method for quantitative PCR detection on CYP2C9 and VKORC1 gene polymorphism. CYP2C9 and VKORC1 gene polymorphism is detected by using a method of a real-time fluorescence quantitative PCR Taqman-MGB probe, and in addition, a detection method of double probes of a wild type and a mutant typeis adopted, so that not only is specificity improved, but also the cost is lowered; and the Taqman-MGB probe is prior to a common probe in aspects such as experiment result accuracy, repeatability, specificity and sensitivity. Therefore, the method for quantitative PCR detection on CYP2C9 and VKORC1 gene polymorphism, which is established by the invention, has the advantages of being accuracy inresult, high in specific sensitivity, free of toxin or pollution, low in cost, rapid and efficient, and the like.

The present invention provides a rapid detection kit for CYP2C9 and VKORC1 genotypes based on a POCT mode. The kit contains a fluorescent quantitative PCR primer and probe for detecting CYP2C9 and VKORC1 genotypes, and a cell lysate, wherein the sequence of the PCR primer is shown as SEQ ID NO.1-2 and SEQ ID NO.5-6, and the sequence of the probe is shown as SEQ ID NO.3-4 and SEQ ID NO.7-8. The kitcan realize real-time detection without DNA purification. A sample can be directly added into a reagent for a PCR reaction, the kit is especially suitable for rapid accurate detection of samples withlow DNA content (such as oral exfoliated cells), and the detection accuracy is 99% or more. The detection sensitivity is high, and the detection limit of genomic DNA as low as 0.125 ng can be accurately detected. The whole detection takes a short time, and a detection result can be obtained within one hour and provides a medicine use basis for a doctor at the first time, thereby reducing the riskof medicine administration.

The invention provides a warfarin medication gene detection kit. The warfarin medication gene detection kit comprises a primer pair used for detecting polymorphism of to-be-detected loci of CYP2C9 * 2, CYP2C9 * 3, CYP4F2 * 3 and VKORC1 genes, probes, an inhibitor, a PCR reaction liquid, a positive quality control product and a negative quality control product; and the to-be-detected loci are at least one of an rs1799853 locus of CYP2C9 * 2 (430C>T) gene, an rs1057910 locus of CYP2C9 * 3 (1075A>C) gene, an rs2108622 locus of the CYP4F2 * 3 (1297G>A) gene and an rs9923231 locusof the VKORC1 (-1639G>A) gene. The invention also provides a use method of the warfarin medication gene detection kit. The problems of high sample treatment requirement, long detection period, complex operation, high cost, low specificity and the like during gene polymorphism are solved.

A probe for detecting a polymorphism at position −1639 of the VKORC1 gene, the probe comprising an oligonucleotide having a nucleotide sequence having a length of 10 to 50 nucleotides, which nucleotide sequence comprises the nucleotides 80 to 89 of SEQ ID NO:1 or 2 and has a homology to SEQ ID NO:1 or 2 except that the nucleotide corresponding to the nucleotide at position 80 in SEQ ID NO:1 or 2 is cytosine, which nucleotide corresponding to the nucleotide at position 80 is labeled with a fluorescent dye.