317 results about "Single-base extension" patented technology

Genetic polymorphisms can change protein structure and function, altering dispositions to diseases and conditions. A single nucleotide polymorphism is the smallest genetic mutation and the most difficult to detect. However, single nucleotide polymorphisms also make up 90% of known genetic mutations, thus identifying such polymorphisms is essential. Single base extension uses the affinity of one base for its complementary base to detect polymorphisms, including single nucleotide polymorphisms. Planar waveguides are used as the platform for single base extension enabling rapid, real time detection of genetic polymorphisms. Detection limits in the picomolar range can be obtained. Signals from the non-matched DNA bases are in the range of the blank signal. Detection times of 5 minutes are reported.

The invention discloses a magnetic separation-based multi-sample multi-site high-flux nucleic acid analysis system which comprises four parts, namely an XYZ three-axis mechanical arm and control circuit, an accurate temperature control system worktable and control circuit, a high-flux fluorescence signal detection worktable and control circuit and system control software. According to the system, a porous plate serves as a reaction container, various function modules are contained to be matched with the system, various biochemical reactions in the nucleic acid detection process can be finished under coordination and control of the system control software, and the specific functions comprise accurate liquid transfer, accurate working plate transfer, rapid magnetic separation, automatic sucker installation and discard, accurate temperature control process, low-temperature sample storage, high-flux fluorescence signal detection and system software overall control. According to the magnetic separation-based multi-sample multi-site high-flux nucleic acid analysis system, a magnetic separation technology, a particle microarray technology, a single base extension technology and an automation system are combined, and a novel high-flux nucleic acid detection system is established on the basis of a series of novel high-flux, automatic, high-specificity and high-practicality single base difference detection methods.

The invention relates to a method for detecting 20 hot mutation sites of four deaf genes such as GJB2, SLC26A4, GJB3 and 12S rRNA in a clinical sample, designing amplimers and single base extension primers of different sites aiming at abnormal SNP gene sites of the four genes by a matrix-assistant laser desorption / ionization flight time mass spectrum detection method and then carrying out mass spectrometry of a specific SNP site genotype.

The invention belongs to the field of forensic medicine genetics and in particular relates to a forensic medicine compound detection kit based on a Y chromosome SNP (single nucleotide polymorphism) genetic marker for individual recognition and genetic relationship identification by a legal medical expert. The forensic medicine compound detection kit provided by the invention is used for carrying out forensic medicine genetic relationship identification and individual recognition on human biology detection materials by utilizing a Y chromosome SNP genetic marker. According to the technical scheme for solving the technical problem, the forensic medicine compound detection kit based on the Y chromosome SNP genetic marker comprises a separated and packaged compound amplification primer mixture, a multiple single-basic-group extension reaction primer mixture, an allele typing standard mixture, a compound amplification reaction mixture and a single-basic-group extension reaction mixture. The kit provided by the invention can be applied to detection of common degradable materials in forensic medicine.

The invention belongs to the field of forensic medicine, and in particular relates to a medicolegal composite assay kit based on twenty triallelic SNP (single nucleotide polymorphism) genetic markers. The kit consists of a composite amplification primer mixture, a multiple single-base extension reaction primer mixture, an allelic typing reference material mixture, a composite amplification reaction mixed liquor and a single-base extension reaction mixed liquor, which are packed in a separate way. The kit can quickly and exactly obtain the typing of twenty triallelic SNP (single nucleotide polymorphism) genetic markers of a biologic assay material at one time to ensure the individual source of the assay material and provide a new technical means for the personal identification by a composite amplification reaction and a multiple single-base extension reaction in a single pipe and a capillary electrophoresis platform generally used in a medicolegal heredity lab. Therefore, the kit has good application prospect in the field of forensic medicine.

The invention relates to a complete-set primer applied to detecting of skin aging related susceptible genes SNP (single-nucleotide polymorphism) and application thereof. The complete-set primer is prepared from amplification primer pairs of 29 SNP sites of 22 skin aging related susceptible genes of human genome DNA and extension primers; a pair of multiple PCR amplification primers and a piece ofsingle-base extension primer are designed for each site respectively, and an MALDI-TOF MS (matrix-assisted laser desorption / ionization time-of-flight mass spectrometry) method is utilized to detect SNP site genotyping of a sample to be detected. The skin aging related susceptible gene detecting method based on gene molecular subtyping disclosed by the invention can finish a plurality of SNP genotyping in the same reaction at the same time, so that accuracy, high efficiency and low cost are achieved. The gene detection disclosed by the invention can be applied to predicting of congenital agingresistance of the skin; thus, a scientific basis is provided for an external skin care scheme.

The invention relates to a genetyping method based on SNP site nucleic acid mass spectrum detection and application thereof. The genetyping method comprises: (1) designing a specific primer with respect to sequence SNP site of a target gene; (2) amplifying a target segment containing the specific SNP site of the target gene utilizing single-pipe multi-PCR; (3) digesting PCR product segment utilizing restriction enzyme; (4) performing single basic group extension reaction; (5) desalting and purifying; and (6) detecting and analyzing the gene sequence of the specific SNP site of the target gene.The genetyping method based on SNP site nucleic acid mass spectrum detection is high in detection accuracy, high in experiment repetition, large in flux, and low in cost. The invention also disclosesapplication of the genetyping method based on SNP site nucleic acid mass spectrum detection in single nucleotide polymorphism. The method can specifically and simply type the target gene or pathogenic microorganism, thereby greatly improving the typing efficiency.

The invention discloses a method for a paternity test through utilizing a specific single nucleotide polymorphism (SNP) mark combination which comprises 60 SNP marks. The nucleotide sequences of the forward primers of the SNP marks are shown as SEQ ID NO.1-60; the nucleotide sequences of downstream primers are shown as SEQ ID NO.61-120; and the nucleotide sequences of single base extension primers are shown as SEQ ID NO.121-180. The method utilizes a flight mass spectrum method, the SNP mark combination is used for human paternity tests, and the method has the advantages of accurate result, quickness and convenience in method and high flux.

The invention discloses a complete-set primer for SNP (Single Nucleotide Polymorphism) sites of drug-metabolizing enzyme genes and application thereof. The complete-set primer comprises amplificationprimers and extension primers of 9 SNP sites of 7 drug-metabolizing enzyme associated genes of human genome DNA, wherein a pair of multiple PCR amplification primers and a single-base extension primerare respectively designed for each site, and an MALDI-TOF MS (Matrix-Assisted Laser Desorption Ionization-Time of Flight Mass Spectrometry) method is adopted to detect typing of the SNP sites of a sample to be detected. The complete-set primer disclosed by the invention has the beneficial effects that a detection method for the drug-metabolizing enzyme genes based on gene molecule typing is established, and the typing of multiple SNPs can be finished at the same time in the same reaction, so that the accurate and efficient effects are achieved and the cost is low; the covered genes and sitesare related to the metabolizing enzyme genes related to 8 major types of common drugs for children, and the coverage is most complete in gene detection products of safe drugs used for children so far.

The invention belongs to the technical field of biology, and particularly relates to a kit for detecting efficient typing of gene loci related to clopidogrel drug resistance reaction. The kit is based on a Sequenom MassARRAY typing system, and comprises PCR (polymerase chain reaction) amplification primers and single-base extension primers of gene loci CYP2C19*2 (rs4244285) and CYP2C19*3 (rs4986893). The sequences of the PCR amplification primers of gene loci are respectively disclosed as SEQ ID NO.1, SEQ ID NO.2, SEQ ID NO.3 and SEQ ID NO.4. The sequences of the single-base extension primers of gene loci are respectively disclosed as SEQ ID NO.5 and SEQ ID NO.6. Before the subject takes clopidogrel, whether the gene loci of the subject are mutated is detected to instruct the individualized reasonable application of the subject, thereby providing references for individualized antiplatelet therapy.

The invention belongs to the technical field of gene detection, and relates to a reagent kit for detecting cardiovascular disease medication genes and a detection method. The reagent kit comprises a detection reagent for 17 mutant sites of 11 genes, in accordance with exceptional gene variation sites of the 11 genes, an amplification primer and a single-basic-group extension primer of each site are designed, through multiplex PCR and a multiple single-basic-group extension reaction, mass spectrometric analysis is performed on a specific product, and the genotype of each site is judged. The detection specificity is high, the sensitivity is high, and the reagent kit can be applied to scientific research, medicine treatment effect research, cardiovascular disease relevant investigation and cardiovascular system clinical medication guidance.

The invention relates to a kit for predicting nasopharynx cancer onset risk, and a corresponding gene chip. The kit comprises a PCR (Polymerase Chain Reaction) amplification primer and a single base extension primer which are used for detecting 7 SNP (Single Nucleotide Polymorphism) loci of personal genome, wherein the 7 SNP loci are rs1412829, rs1572072, rs28421666, rs2860580, rs2894207, rs6774494 and rs9510787; the kit further comprises a PCR amplification primer and a single basic extension primer which are used for detecting 4 SNP loci of personal genome, wherein the 4 SNP loci are rs2853668, rs31489, rs402710 and rs4635969; the kit further comprises a nested PCR amplification primer for detecting a single nucleotide polymorphism locus G155391A with EB (Elzatein-Barn) virus subtype specificity related to nasopharynx cancer. The gene chip comprises upstream and downstream primers and probes for detecting 11 SNP loci of personal genome, and upstream and downstream primers and probes for detecting a single nucleotide polymorphism locus G155391A with the EB virus subtype specificity related to nasopharynx cancer, wherein the 11 SNP loci are rs1412829, rs1572072, rs28421666, rs2860580, rs2894207, rs6774494, rs9510787, rs2853668, rs31489, rs402710 and rs4635969.

A method for detecting variation of gene based on fluorescence quenching quantification comprises: performing single base extension at a specific site of a gene to be detected with marked probes and marked dideoxyribonucleotide triphosphate; detecting fluorescence quenching values, and determining SNP of the gene to be detected. The present invention also provides an oligonucleotide probe for the method.

The invention discloses an automation micro-flow workstation and a method for carrying out a solid phase PCR reaction. The method refers to a method for realizing the solid phase PCR reaction based on a micro-fluidic chip. The workstation comprises at least one micro-fluidic chip, a fluid path system, a temperature control device and a control system, wherein each chip comprises one or more independent reaction channels; each reaction channel comprises an inlet, an outlet, one or more reaction chambers and / or flow channels connected in series or in parallel; an oligonucleotide sequence is anchored to the bottom surface of each reaction chamber so as to perform solid phase PCR amplification and / or solid phase single base extension; the fluid path system controls to introduce different reaction reagent solutions so as to flow to corresponding reaction chambers; the temperature control device is used for controlling and regulating the temperature in the reaction chambers; and the control system is connected with the fluid path system and the temperature control device and is used for controlling the solid phase PCR amplification and / or solid phase single base extension performed in the reaction chambers of the micro-fluidic chips in the fluid path system and the temperature control device, and the reactions comprise a denaturation-elution reaction and an elution-denaturation reaction. The workstation has the advantages of one-stop automation, high flux, low cost, convenience in operation, rapidness, high efficiency and high accuracy.

The invention belongs to the field of forensic medicine genetics and in particular relates to a forensic medicine compound detection kit based on a Y chromosome SNP (single nucleotide polymorphism) genetic marker for individual recognition and genetic relationship identification by a legal medical expert. The forensic medicine compound detection kit provided by the invention is used for carrying out forensic medicine genetic relationship identification and individual recognition on human biology detection materials by utilizing a Y chromosome SNP genetic marker. According to the technical scheme for solving the technical problem, the forensic medicine compound detection kit based on the Y chromosome SNP genetic marker comprises a separated and packaged compound amplification primer mixture, a multiple single-basic-group extension reaction primer mixture, an allele typing standard mixture, a compound amplification reaction mixture and a single-basic-group extension reaction mixture. The kit provided by the invention can be applied to detection of common degradable materials in forensic medicine.

The invention relates to PCR amplimers, single base extension primers, a detection kit and a detection method for the polymorphism of the human IL28B gene. The kit comprises the PCR amplimers and the single base extension primer designed specific to the polymorphism of the site rs12979860 (T / C), and the PCR amplimers and the single base extension primer designed specific to the polymorphism of the site rs8099917 (T / G). The above-mentioned primers are used for PCR amplification, SAP reaction and single base extension, and then mass spectrometry is employed for analysis of the forms of the polymorphism of the above-mentioned sites. The detection method for the polymorphism of the human IL28B gene can rapidly, specifically and highly efficiently detect the polymorphism of the two above-mentioned SNP sites of the IL28 gene; thus, prediction can be carried out on the curative effects of a standard hepatitis C therapy scheme, differentiated dealing and treatment can be carried out on patients with different types of hepatitis C, and the whole diagnosis and treatment level of hepatitis C is improved.

The invention discloses a flight mass spectrum biochip for health risk assessment. The biochip includes a sample analyte and a mass spectrum chip matrix that undergo cocrystallization. The sample analyte comprises a?target?gene, a PCR primer and a single base extension primer used for innate potential assessment. The invention also discloses a detection method making use of the chip. The method consists of: extracting the sample DNA of a detection subject; designing a PCR primer and a single base extension primer; conducting target gene PCR amplification and single base extension on the sample DNA to form a sample analyte; and subjecting the sample analyte and the mass spectrum chip?matrix?to cocrystallization, and detecting the SNP locus of the target gene. The gene chip provided in the invention can reveal the risk of a detection subject in suffering from a disease in the aspect of heredity so as to further provide personalized health intervention measures, thus enabling the detection subject to maintain a good health level.

The invention relates to a multiple PCR (polymerase chain reaction) primer combination used for a human paternity test. Eighty SNP (single nucleotide polymorphellosm) genetic markers can be detected by virtue of the primer combination. Nucleotide sequences of primers at middle and upper reaches in the primer combination are respectively shown in SEQ ID NO.1-80; nucleotide sequences of primers at a lower reach in the primer combination are respectively shown in SEQ ID NO.81-160; and nucleotide sequences of single-base extension primers are sequentially shown in SEQ ID NO.161-240. According to the multiple PCR primer combination, the human paternity test is carried out by preferably utilizing a flight time spectral method, a result is accurate, a method is rapid and simple, and flux is high, so that the multiple PCR primer combination has a good application prospect.

The invention discloses a primer combination for detecting skin anti-aging capability genes. The skin anti-aging capability genes comprises collagen multiplication capability gene COL1A1, a skin moisture retaining and keeping capability gene AQP3, a radiation protection capability gene ASIP, ultraviolet damage repair capability genes ERCC2 and LOC105374069, a skin detoxifying capability gene GSTP1, a skin sensitivity gene IL6R and an estrogen level gene DIAPH2, and corresponding PCR primers and single-base extension reaction primers are designed aiming at the eight genes. The invention further discloses a method for performing gene detection by utilizing the primer combination. The method comprises the following steps: carrying out sample DNA extraction, PCR amplification and single-base extension and nucleic acid mass spectrometric analysis to obtain corresponding genotypes, and then carrying out analysis and scoring, thereby scientifically evaluating the difference of individual skin aging processes, providing scientific and precise recommendations for daily skin care and life habits of individuals, and also being helpful to reasonable selection of skincare products of individuals according to skin types.

The invention relates to a kit for detecting susceptibility gene SNP locus of nasopharynx cancer, which is realized through the steps as follows: detecting a PCR amplimer and a single-base extension primer of an SNP locus rs2860580 of a susceptibility gene CDKN2A-CDKN2B of nasopharynx cancer; detecting PCR amplimers and single-base extension primers of SNP loci rs1572072 and rs9510787 of a susceptibility gene TNFRSF19 area of nasopharynx cancer; detecting PCR amplimers and single-base extension primers of SNP loci rs28421666,rs2860580 and rs2894207 of a susceptibility gene HLA of nasopharynx cancer; and detecting a PCR amplimer and a single-base extension primer of an SNP locus rs6774494 of a susceptibility gene MDS1-EVI1 of nasopharynx cancer. The kit provided by the invention can be used for simultaneously detecting the SNP loci of a plurality of susceptibility genes of nasopharynx cancer, and provides reference for estimating risk degree of individuals affected by nasopharynx cancer, general survey of populations with high nasopharynx cancer incidence, screening of high risk group attacked by nasopharynx cancer and implementation of relative precautionary measures.

The invention belongs to the field of biotechnology, and relates to a method and a kit for identifying free DNAs in serum by adopting a mass spectrometer and applications of the method and the kit. The method comprises the steps of PCR reaction, PCR product purification, single base extension, extension product purification and detection by adopting the mass spectrometer; the kit comprises a PCR reaction reagent, a purifying reagent and a single base extension reaction reagent. Compared with the prior art, the method and the kit provided by the invention have the advantages of high sensitivity, strong specificity, simplicity and safety, high speed, high flux, and the like, and a relatively large amount of time is saved.

The invention provides a primer combination for detecting gene polymorphic sites relevant to clopidogrel resistance, wherein the sites comprise the polymorphic sites including CYP2C19*2, *3, *4, *5, *17 and the polymorphic site 3435C>T of ABCB1. According to the scheme, multiple PCR primers are utilized for amplifying gene segments where relevant gene loci are located, after the amplification product is treated, single base extension is carried out on the to-be-detected loci, then molecular weight difference detection is carried out on the extension product by adopting a flight time mass spectrum, through data analysis, the result of the relevant gene for the clopidogrel resistance of a patient is rapidly detected. The invention further provides a detection product prepared by adopting the primer combination and applications of the detection product, and thus references are provided for the individualized medication of clopidogrel.

The invention provides a primer group capable of simultaneously detecting 27 respiratory tract pathogens based on a nucleic acid mass spectrometry technology, a kit and application, and belongs to thefield of pathogen nucleic acid detection. According to the kit, PCR amplification primers and single-base extension primers are designed for 27 clinically common respiratory tract pathogens. Pathogenic bacteria corresponding primer and single base extension primer sequences are shown in SEQ ID NO: 1-81. According to the nucleic acid mass spectrometry, a multiple RT-PCR technology and a single base extension technology are integrated, pathogen target genes are amplified through multiple RT-PCR, and the detection sensitivity is improved; in the single-base extension reaction, one base amplification is carried out on an extension primer, and pathogen identification is carried out by distinguishing the molecular mass of an extension product, so that the specificity is high. Meanwhile, the flux of the nucleic acid mass spectrum is high, amplification of 40 genes can be achieved at most through one reaction hole, and 384 samples can be analyzed at most at a time by means of the chip technology; the kit has the advantages of high automation degree, high detection flux, high sensitivity and good specificity.

The invention discloses a flight mass spectrum biochip for innate potential assessment. The biochip includes a sample analyte and a mass spectrum chip matrix that undergo cocrystallization. The sample analyte comprises a target gene, a PCR primer and a single base extension primer used for innate potential assessment. The invention also discloses a detection method through the chip. The method consists of: extracting the sample DNA of a detection subject; designing a PCR primer and a single base extension primer; conducting target gene PCR amplification and single base extension on the sample DNA to form a sample analyte; and subjecting the sample analyte and the mass spectrum chip matrix to cocrystallization, and detecting the SNP locus of the target gene. The gene chip provided in the invention can get an in-depth understanding of the potential ability of a person, make assessment on individual innate potential, and forecast the problems a person may encounter in the growth process in advance, thus avoiding receiving education blindly. Therefore, the biochip provided in the invention can help the detection subject to make targeted life career planning.

The invention discloses a gene detection product for genetic deafness, and particularly relates to a reagent kit for detecting multiple deafness genes at the same time. The reagent kit uses multiple PCR amplification primers and single base extension primers to simultaneously detect multiple hotspot mutations in the same tube, and has the advantages of quick diagnosis, easy operation and low cost. The invention also discloses a method for combining multiple PCR- single base extension-mass spectrum, and the method can be used to detect the genotype of the deafness gene accurately, sensitively and in high throughput, thereby being helpful for the popularization of deafness gene screening.

The invention provides a primer set for amplifying genes related to exercise ability and a detection kit, and relates to the technical field of application of SNPs. The primer set comprises a specificprimer and a single base extension primer; the nucleotide sequence of the specific primer is shown in SEQ ID NO. 1 to 52; and the nucleotide sequence of the single base extension primer is shown in SEQ ID NO.53 to 78 n. The amplification primer and extension primer designed by the invention can detect 26 SNP sites in the same reaction, and are used for developing exercise ability detection kit and guiding reasonable movement. In terms of exercise guidance, the genetic test of the invention can understand the endurance, explosive power, strength and the like of the human body, and provide scientific reference for rational exercise. The movement-related genes are jointly detected and all the items are detected at a time, so that the primer set and the detection kit are most comprehensive incoverage of detection genes in the exercise ability gene detection products.

The invention belongs to the technical field of forensic medicine, and particularly relates to a 26 mitochondria SNP genetic markers-based forensic medical composite detection kit which is used for carrying out forensic medical personal identification on the unknown human biological samples by utilizing SNP genetic markers. The 26 mitochondria SNP genetic markers-based forensic medical composite detection kit comprises a multiplex amplification primer mixture, a multiple single base extension reactant mixture and an allele parting standard mixture, wherein the mixtures are separately packaged. According to the kit, the technologies of complex amplification in single tubes and multiple single base extension are adopted, so that the gene parting of the 26 mitochondria SNP genetic markers of the biological samples can be obtained at one time and the forensic medical personal identification can be rapidly carried out.

The invention relates to a method and kit for detecting human alcohol metabolizing capacity gene mutation sites. The detection method comprises the following steps: designing specific primers by aiming at an alcohol dehydrogenase ADH1B gene rs1229984 site and an acetaldehyde dehydrogenase ALDH2 gene rs671 site, and performing specific polymerase chain reaction (PCR) amplification so as to obtain target fragments; and then performing enzyme disgestion, single base extension and desalting purification treatment, and then detecting analytic sequences by use of a nucleic acid velocitron. The method for detecting the human alcohol metabolizing capacity gene mutation sites is high in detection accuracy, high in experiment repeatability, great in flux and low in cost. The invention also providesa kit for detecting human alcohol metabolizing capacity gene mutation sites. The kit comprises a specific primer pair for amplifying an alcohol dehydrogenase ADH1B gene rs1229984 site gene and an acetaldehyde dehydrogenase ALDH2 gene rs671 site gene. The kit for detecting human alcohol metabolizing capacity gene mutation sites is capable of simplifying experimental procedures and has the advantages of short manual operation time, low difficulty and high experiment automation degree.

The invention discloses a primer combination sequence and kit for detecting child safe medication-related gene mutation sites. The kit includes 23 pairs of amplification primers and 23 single-base extension primers; the 23 pairs of amplification primers can specifically amplify the 23 common mutation site regions of child safe medication-related genes, and the 23 single-base extension primers areused to detect the 23 mutation site genotypes of the child safe medication-related genes; and the kit further includes special reagents for pretreatment and detection. The kit provided by the invention can realize one-hole detection of the 23 mutations related to the clinical child safe medication-related genes, has high sensitivity, strong specificity and high accuracy, and is simple to operate,low in cost and high in throughput, the detection is fast, automatic interpretation of results is realized, and the clinical promotion and application are easy to realize; and the kit can be applied to the detection of the genes related to safe medication of newborns or children, guide clinical correct medication, avoid medication risks, reduce medication injury, and truly achieve precise medication.