313 results about "Mutation type" patented technology

The invention discloses an artificial fixed-point rice dense and erect panicle (DEP1) gene mutant body and application thereof. The rice DEP1 gene mutant body as well as a corresponding allele is obtained by modifying a 5th exon of a DEP1 gene by adopting a CRISPR / Cas9 gene targeting modification technology, so that a base of the 5th exon is replaced, lost and inserted. The artificial fixed-point rice DEP1 gene mutant body and the application thereof disclosed by the invention have the benefits that the artificial mutation is performed on the 5th exon of the rice DEP1 gene by adopting the CRISPR / Cas9, and the mutant body capable of obviously improving the rice yield and the allele are screened; the DEP1 gene mutant body disclosed by the invention as well as the corresponding allele can improve the yield of rice plants by 13-51 percent and is even superior to a natural mutation type.

The invention discloses a probe and a sequence combination for simultaneous detection of various mutation types and particularly provides a kit, a method, a gene chip, a probe and a sequence combination which are high in sensitivity and specificity and used for simultaneous detection of point mutation, short segment insertion and deletion, copy number variation and fusion genes of 57 tumor driver genes listed in a detection list. The invention further discloses a sequence combination and probe associated gene chip and kit and a method for simultaneous detection of various mutation types. By detection of body fluid including blood, hydrothorax, abdominal dropsy and the like and tumor frozen tissues or paraffin sections, comprehensive tumor driver gene mutation information can be acquired, sensitivity and accuracy in tumor gene detection are improved, and accordingly clinicians can be assisted in making of individualized medication schemes to achieve best accurate treatment effects.

The invention relates to a disease risk predication method and system. The method comprises the following steps of: S1, receiving input gene sequencing result information; analyzing the input gene sequencing result information to obtain all mutant gene information; S2, looking up disease gene mutant information of corresponding gene sites from a precision medical knowledge base according to the gene sites corresponding to the mutant gene information, so as to obtain at least one piece of disease gene mutant information; S3, matching each piece of the disease gene mutant information with all the disease gene mutant information to obtain the similarity of each piece of the mutant gene information, and all the disease gene mutant information; and S4, obtaining a risk predication table aiming at the gene sequencing result information according to the similarity of each piece of the mutant gene information, and all the disease gene mutant information. By analyzing an association relation of susceptibility genes, mutation sites, mutation types and diseases, disease risk predication is carried out on healthy people and general disease risk grade predication is provided; disease risks in the future are reduced.

The invention provides a method and device for detecting single sample body cell mutation sites in abnormal tissue and a storage medium. The method includes the following steps that the effective sequencing sequence of an abnormal sample and the effective sequencing sequence of a simulated normal sample are obtained; in the effective sequencing sequences, basic groups, different from basic groups of the simulated normal sample, of the abnormal sample are obtained, according to the mutation basic group frequency, the types of the basic groups of the abnormal sample and the simulated normal sample are judged, the FISHER is then used for accurately detecting the difference of the types of the basic groups, and according to the difference, the mutation type is judged; by filtering the mutation types, false positive mutation and germ cell mutation are removed, and high-reliability somatic cell mutation sites are obtained. The method has the advantages of being high in sensitivity and specificity, has high sensitivity in mutation detection of known mutation genes, and new mutation genes can be found.

The invention relates to a kit, a method and an application for detecting mutation of predetermined locus in DNA sample. The method for detecting mutation of predetermined locus in DNA sample comprises the following steps: using an amplification primer for performing PCR amplification reaction on the DNA sample, using an extended primer and dd NTP; using the amplification product as template, performing the oligonucleotide extension reaction, so as to obtain the extended primer having a 3' end connected with a base, which is the extended product; and based on the molecular weight of the extended primer, determining the mutation type of the predetermined locus. The invention can effectively determine whether there is mutation in the predetermined locus and can determine the mutation type.

The invention discloses a tumor cloning mutation detection method and device based on next-generation sequencing and a memory medium. The method provided by the invention comprises the steps of carrying out mutation detection on a comparison file of paired tumor and normal samples through utilization of mutation detection software, computing a mutation frequency, and selecting segments with high sequencing quality as a statistics result; carrying out copy number and purity detection on the paired tumor and normal samples through utilization of purity detection software; combining small segments into big segments, and annotating the copy number in a mutation area; and computing a proportion of the mutation in a tested tumor tissue through utilization of a beta distribution model according to tumor sample purity and copy number detection results, thereby judging a tumor cloning mutation type. According to the method provide by the invention, influences of the sample purity and multiploidon the detection are avoided, the mutation type detection is relatively accurate, the subcloning mutation with clinical significance can be effectively identified, and the foundation for accurately and deeply researching a tumor cloning evolution process and searching a tumor therapy molecular mechanism is laid.

The invention provides primers, a method and a chip for detecting alpha and / or beta-thalassemia point mutation and deletion mutation based on whole-gene capture sequencing and application of such primers, such method and such chip. The primers, the method, the chip and application thereof have the advantages that through designing of capture probes, relevant genes involved in alpha-thalassemia and beta-thalassemia are enriched and all mutation information including SNP and indel in full-length sequences of genes is detected; through addition of autosome, X-chromosome and Y-chromosome regions as well as upstream and downstream regions of coded genes as references, structure variations such as SNV and CNV are detected; compared with existing various hotspot mutation site detection technologies, the method is capable of detecting hotspot mutation information as well as some rare mutations and undiscovered new mutation types to detect and analyze full-length sequence specificity of target genes, fully covers the mutation types and makes up the defect that a conventional detection method easily causes missing detection of low-frequency mutations and rare mutations greatly.

The invention relates to a gene mutation type recombined protease K and an industrialized production method thereof. Specifically, the invention relates to the gene mutation type recombined protease K which is obtained by performing gene reformation and protein engineering and has an enzymatic activity being more than two times of the enzymatic activity of the same natural protein, and further relates to the industrialized production method and a technical process for utilizing yeast cells to large-scale culture expressing foreign proteins and prepare the gene mutation type recombined protease K.

The invention relates to a method for identifying gene mutation types in the field of gene analysis as well as a special chip and a kit thereof. The gene mutation detecting method comprises the following steps: taking a genome to be detected from a human tissue as a template, carrying out multiple allele special PCR amplification by a primer group that is designed aiming at special mutant sites and DNA polymerase without 3'-5' end exonclease activity, then hybridizing the obtained PCR product and an oligonucleotide probe (allele special probe) on the gene chip, and confirming mutation types of all gene sites according to the hybridizing result. The allele special probe is designed aiming at special gene types of gene mutant sites to be detected. The invention can detect gene mutations in comprehensive, systemic and high-flux ways and has light environmental pollution as well as simple and rapid operation compared with PCR-RFLP and a sequencing method.

The invention relates to 86 types of particular mutation type atlases of SLC26A4 gene related to hearing loss in Chinese crowd, 27 types of relative hot-spot mutation type atlases and frequency information thereof, 13 types of hotspot mutation type atlases and frequency information thereof, and 2 types of hottest-spot mutation type atlases and frequency information thereof. 59 SLC26A4 gene mutation types are newly discovered in the Chinese crowd, wherein, 47 mutation types lead to the change of encoded protein amino acid of the SLC26A4 gene or influence genetic transcription and translation, 6 mutation types lead to base change rather than the change of amino acid, and 6 types are intron mutation types of the SLC26A4 gene. The discovery has a vital practical significance in developing an SLC26A4 gene diagnosing chip and a kit, which conform to hereditary features of the Chinese crowd suffering hearing loss.

The invention relates to a gene detection kit, in particular to a helicobacter pylori (HP) type and drug-resistant mutation gene detection kit. The kit comprises PCR (polymerase chain reaction) solutions and nucleic acid membrane strips for HP type and drug-resistant mutation gene detection; the PCR solutions comprise a PCR solution I, a PCR solution II, a PCR solution III and a PCR solution IV; the PCR solutions also contain a pair of internal control primers respectively. The HP type and drug-resistant mutation gene detection kit can be used for distinguishing two types of HP in one test, and can be used for detecting 15 mutation types of 9 hot spot mutational loci related with drug fastness of 5 therapeutics, a VacA gene and a CagA gene are used for typing, and a reference basis is provided for judgment of illnesses. Mutation types of drug-resistant mutation detection are more and more comprehensive, and the condition of genotypic resistance of HP from which a patient suffers is quickly and comprehensively evaluated.

The invention discloses a primer group and a method for detecting BRCA1 / 2 gene mutation. The primer group comprises three primer pools, 97 pairs of primers in total, and the sequences of the upstreamand downstream primers are as shown in SEQ ID NO.1-194. The method comprises the following steps: first, extracting DNA of a to-be-detected sample, carrying out multiplex PCR amplification by adoptingthe three primer pools, and combining target fragments obtained through amplification by adopting the three primer pools, thus constructing a library, and finally, carrying out library sequencing andbiological information analysis, thus obtaining the BRCA1 / 2 gene mutation condition of the sample. The method has the advantages of high flux, high accuracy, high sensitivity, high automation degree,low demanded quantity of samples, low cost, rapidness, detection for various mutation types, simultaneous detection for multi-site mutation and the like, can be applied to tumor high-risk screening,medication guidance, prognosis and the like, has great significance in domestic BRCA1 / 2 gene mutation detection and pathogenicity analysis and evaluation, and has a wide application prospect.

The invention discloses a somatic mutation site excavation method based on genomic sequencing. By means of depth re-sequencing on various tissue of a wild type shaddock and a mutation type shaddock, the obtained high-quality sequencing reads and the sweet orange genome are compared, and the single nucleotide polymorphism (SNP) of various sets of materials is obtained. Based on SNP sites distributed on some whole genomes, the case-control type correlation analysis is designed on the wild type shaddock and the mutation type shaddock, potential sites related to mutation are obtained and annotated, the chromosome area where the character correlation variation happens is speculated, finally, the potential sites are verified through a Sanger sequencing method, and the variation site of the mutation type shaddock is obtained. The method has the biggest advantages of being low in cost and short in period. The cost for finishing excavating of one mutant mutation site is about 20 thousand, the excavation can be finished in three months, and combining and innovation exploring of the genome advanced technology and the traditional citrus breeding study are expressed. The invention further discloses development and application of the method.

The invention provides a device for detecting somatic mutation. The device comprises an acquisition module, an annotation module, a screening module, a computing module, a mutation type primary judgment module and a mutation type correction module, wherein the screening module comprises a virtual comparison set, and the virtual comparison set contains 561 cases of mutation information of leukocytes. According to the device, detection data is compared with a database including the virtual comparison set through the screening module, so that embryonal system mutation sites are screened; the computing module is utilized to obtain a mean value and a standard deviation of embryonal system mutation frequency of all chromosomes through computing; then the mutation type primary judgment module and the mutation type correction module are utilized to analyze mutation frequency features of all different mutation sites to determine the type of the mutation sites; and the primarily determined mutation type is corrected through the database of the mutation sites of the known mutation type, so that the mutation sites with somatic mutation in samples are screened out. Through the device, detection accuracy of the mutation sites with somatic mutation is improved.

The invention relates to a method for detecting human beta-globin gene mutation, in particular to a method for detecting the beta-globin gene mutation by a modified molecular beacon melting curve. The method comprises the following steps of: designing and preparing a corresponding modified molecular beacon in region of a beta-globin gene needing mutation detection; designing an upstream primer and a downstream primer on the periphery of the designed modified molecule beacon, and performing PCR amplification on fragments containing the region to be detected by using the upstream primer and the downstream primer; and after PCR amplification is finished, analyzing the melting curve, and judging whether a nucleotide sequence to be detected has the gene mutation and possible mutation types according to the change of the melting point of the modified molecular beacon.

The invention provides a microsatellite unstable site screening and analysis model construction method and device. The method comprises the following steps of: S1, taking a microsatellite site of a reference genome as a candidate MS site, S2, selecting a plurality of samples with known microsatellite instability detection state information as training set samples, carrying out mutation detection on candidate MS site areas of the training set samples, respectively recording mutation types and numbers, and calculating information entropy of mutation of each candidate MS site, and S3, correlatingthe microsatellite instability detection state information with the entropy of the information entropy of the mutation of each candidate MS site, selecting an entropy threshold of each candidate MS site for distinguishing the microsatellite instability detection result, and screening out the candidate MS site with a high distinguishing degree as an MSI site. According to the scheme, the problemsof low detection efficiency caused by excessive fixed MSI sites and low detection specificity and sensitivity caused by incapability of covering all types of samples in the prior art are solved.

The invention discloses a method for identifying HBV gene mutation type and a chip and a reagent box which are especially used with the method. The method for identifying the HBV gene mutation type takes the detected the genome of the hepatitis B virus as the template and uses the following primer group and a DNA polymerase which has no exonuclease activity from the 3'end to 5' end for implementing multiple PCR magnification; the primer group comprises a universal primer and a or more than one special primers; the special primer 5' end is a bar code sequence and the length of the bar code sequence is 5 to 25 nt; the bar code sequence has a comparatively low homology with the hepatitis B virus; the special primer totally matches with the corresponding segment of the hepatitis B virus comprising wild base group or mutant base group at a mutation point; the gene chip comprises a plurality of probes and the probes are corresponding to the bar code sequence one to one and every probe only contains one bar code sequence.

The invention relates to the fields of biotechnologies and medicines, particularly discloses a detection primer for EML4-ALK (anaplastic lymphoma kinase) fused gene mutation, a probe and a detection kit, and aims to detect the EML4-ALK fused gene mutation. The detection primer comprises sequences as shown in SEQ ID NO: 1 to SEQ ID NO: 14. The sequence of the probe is as shown in SEQ ID NO: 15, of which the end 5' is modified with FAM or VIC (vasoactive intestinal contractor), and the end 3' is modified with NFQ-MGB (non-fluorescent quencher-myohemoglobin). The kit comprises the detection primer, the probe and a quality control system. The detection primer can specifically identify 13 EML4-ALK fusion mutation types, is high in sensitivity and can detect 5-copied mutations; the whole reverse transcription PCR (polymerase chain reaction) and fluorescence PCR detection process can be completed only by 80 minutes.

The invention discloses a primer probe composition and kit for detecting the polymorphism of a human CYP2C19 gene and application. The primer probe composition comprises three pairs of specific primers for amplifying CYP2C19*2, CYP2C19*3 and CYP2C19*17 sites and three specific fluorescent probes. The primers and the probes have high sensitivity, strong specificity and strong anti-interference capability; the polymorphism of the gene is detected by adopting a manner combining asymmetric PCR (Polymerase Chain Reaction) amplification and a fluorescent probe melting curve analysis technology; different gene types can be effectively distinguished according to the quantity of melting peaks and Tm value and result interpretation is convenient, clear and objective. Single-tube sampling can be usedfor detecting 6 mutation types of 3 gene sites at the same time; the operation is simple and convenient and the detection efficiency is improved; a large batch of samples can be detected and clinicaloperation is facilitated.

The invention discloses a thalassemia gene detection method based on a high-throughput sequencing technology. The thalassemia gene detection method mainly comprises the following steps that alpha & beta-thalassemia related gene fragments are specifically amplified based on PCR amplification primers of a span breakpoint and a mutation site designed by adopting a Gap-PCR method, library establishment is conducted on PCR products, library products are subjected to high-throughput sequencing, sequencing data uses a human genome hg19 as a reference sequence, sequencing depth values of gene loci in a target area chr16: 215000-236000 are analyzed according to a sequence alignment score Q10, and alpha-thalassemia deletion types are determined according to the distribution of the sequencing depth values of different loci; meanwhile sequence alignment is performed through the target area chr16: 215000-236000 and a target area chr16: 5246400-5248600, and alpha & beta-thalassemia mutation types are determined according to the basic types of specific sites. By the adoption of the thalassemia gene detection method, simultaneous detection of 6 mutation genetypes of alpha-thalassemia and 26 mutation genetypes of beta-thalassemia can be achieved.

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 kit for synchronously detecting 12 mutation hotspots of the Chinese population phenylketonuria PAH (phenylalanine hydroxylase) gene. In the kit, 12 loca on the Chinese population PAH gene are used as detection objects; an amplification primer and an extension primer are respectively designed for the mutation type of each locus; each destination section is subjected to PCR amplification and mark extension at the same time; and the gene types of the 12 mutation hotspots are obtained at the same time through capillary electrophoresis analysis. The invention provides a phenylketonuria PAH gene mutation screening kit which is simple, has the advantages of high flux, high performance, low cost and high detection accuracy, is suitable for the Chinese people and suitable for the group screening of the Chinese population phenylketonuria.

The invention belongs to the field of medicines and biology, and relates to a blood plasma cfDNA detection technology-based noninvasive pancreas cancer polygene detection method and kit. The blood plasma cfDNA detection technology-based noninvasive pancreas cancer polygene detection method comprises the following steps: collecting blood, separating blood plasma, and purifying cfDNA; designing the panel of a pancreas cancer characteristic hotspot mutation region; and carrying out PCR primer amplification, constructing a library, and carrying out high-flux next generation sequencing and data analysis. The dynamic change of the mutation type and the mutation frequency of pancreas cancer characteristics in the blood plasma cfDNA is detected according to the treatment process and the pathogenesis development of every individual, so information is provided for pathogenesis monitoring and resistance drugs of the pancreas cancer.

The invention discloses a nested asymmetric PCR reagent kit for detecting alpha 2 globin gene point mutation. The reagent kit comprises a nested asymmetric PCR amplification primer, a fluorescent probe, a quenching probe, an LADNA polymerase and the like. The alpha-thalassemia point mutation genotype of a detected sample is detected by virtue of specific amplification of five point mutation destination fragments WS, QS, CS, CD30 and CD31 of the human alpha 2 globin gene and molecular hybridization and unwinding analysis of a specific fluorescent probe. The reagent kit has good sensitivity and accuracy to wild type and mutation type of alpha-thalassemia five point mutation positions WS, QS, CS, CD30 and CD31, has good repeatability and stability, and is totally suitable for clinical detection of alpha-thalassemia point mutation.

This invention belongs to the medicine curative effect detection technique field; involve a kind of tumor relevant gene mutation PCR detection method and reagent system concretely. Further, involve TaqMan - MGB real-time fluorescence quantitative PCR method of EGFR and ras gene mutation correlated with curative effect of molecule target anticancer drug and reagent system. It mainly includes clicking in the specific sudden change location where the molecule target is correlated with the curative effect of anticancer drug, the peculiar oligonucleotides designed specially primer series and probe series, determine procedure method , reagent system and use. The reagent system mean includes the any peculiar oligonucleotides primer series and real-time fluorescence quantitative PCR reagent systemof the probe.

The invention provides a kit and method for detecting the mutation of an EGFR gene and particularly discloses a method, a primer and a probe for detecting the mutation of the EGFR gene, a kit including prime probe mixing liquid, a primer and a probe for detecting the mutation of a C797S locus, a primer and a probe for detecting the mutation of a T790M locus, a primer and a probe for detecting themutation of a 19del locus and a primer and a probe for detecting the mutation of a L858 locus. According to the method provided by the invention, 0.1% mutation rate can be detected based on a digitalPCR platform; and the method has the advantages that the optimization process is simple, much detected mutation types can be detected, the absolute quantification is realized, the sensitivity is high,samples are easily acquired, and the like.

The invention discloses a primer and probes used for full RAS mutation detection. The primer and probes comprise a full RAS specific primer and the probes, and the full RAS specific primer and the probes comprise twenty seven mutation types of KRAS 2,3,4 exons and twenty three mutation types of NRAS 2,3,4 exons. A detection kit used for full RAS mutation detection extracts a DNA sample and performs a PCR reaction. The specific MGB Blocker probe, the NRAS specific probe and the KRAS specific probe and the primer are adopted and can detect RAS mutation in DNA in tumor tissue; MGB Blocker is used for retarding nonspecific augmentation of NRAS and KRAS, and sensitivity is improved and higher than 1% of ARMS; RAS mutation can be detected in circular tumor DNA. The primer and the probes are easy to operate, low in detection cost, capable of being popularized in a large-scale mode and high in detection speed, and the detection process can be finished after about 2 hours.

The invention relates to a method for detecting multiple mutation types of a genome. A BAM file is used as an input file, a tumor tissue sample is detected, multiple mutation information of differentsample types is detected, and detection of the tumor tissue sample comprises the steps as follows: 1) detection of mononucleotide mutation; 2) detection of insertion and deletion; 3) detection of structural variants; 4) detection of complex variants. According to the method for detecting multiple mutation types of the genome, insertion and deletion are detected with a GeneReader algorithm obtainedthrough combination of a supervised method and an unsupervised method, the local comparison precision is optimized, linear growth with increase of the sequencing depth is realized in the aspect of speed, the detection effect is good, the detection result is accurate, and actual application demands can be well met.

The invention discloses a method for detecting related gene mutations of diabetes drug treatment as well as a special chip and a kit thereof. The method comprises the following steps: taking a genome to be detected from a human tissue as a template, carrying out multiple PCR amplification by a primer group that is designed aiming at special mutant sites and DNA polymerase without 3'-5' end exonclease activity, then hybridizing the obtained PCR product and an allele specific probe on the gene chip, and confirming mutation types of related genes of diabetes drug treatment according to the hybridizing result. The invention can detect known gene mutations closely related to the individual difference of anti-tumor medicine reaction in comprehensive, systemic and high-flux ways and greatly improve the accuracy and the specificity of chip detection.