85 results about "Hotspot mutation" patented technology

The invention discloses a fluorescence detection kit capable of detecting 17 non-syndromic hereditary hearing loss susceptibility genes. The kit adopts 17 pairs of specific primers to conduct genetic typing on the hearing loss susceptibility genes, and 17 hotspot mutations in four most common Chinese hearing loss related genes can be detected at the same time in a single tube within 3 hours. The kit comprises primer combinations of 17 polymorphic sites of hereditary hearing loss on a GJB2 (CX26) gene, an SLC26A4 (PDS) gene, a GJB3 gene and a 12SrRNA (MTRNR1) gene, can be used for accurately judging the wild type, the pure mutant type or the hybrid type of the 17 sites, and achieves diagnosis and screening of the hearing loss genes. The kit provided by the invention can be applied to rapidly and efficiently detecting the hearing loss genes and is a rapid, convenient, economical and efficient screening kit for hearing loss virulence genes.

The invention discloses a method and a kit for detecting a non-small cell lung cancer drive gene mutation spectrum, and an application. The method comprises the following steps of: designing 15 pairs of amplification primers for amplifying the exon segments of the seven related genes of non-small cell lung cancer, dividing the amplification primers into 6 groups, and preparing an amplification primer mixed solution, performing multiple PCR (polymerase chain reaction) amplification on the to-be-detected samples by the amplification primer mixed solution respectively, and then performing enzymatic digestion; and designing 39 extension primers used for detecting hotspot mutation sites, dividing the extension primers into 6 groups corresponding to the amplification primer mixed solution, and preparing an extension primer mixed solution, performing extension reaction on the digested PCR product, then performing enzymatic digestion, performing capillary electrophoresis on the obtained product, and making a result judgment via software analysis. The kit provided by the invention comprises the amplification primers for amplifying the exon segments of the seven related genes of non-small cell lung cancer, and the extension primers used for detecting hotspot mutation sites. The method and the kit provided by the invention are simple, high in flux, and short in time consumption.

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 discloses composition for detecting colorectal cancer hotspot gene mutation sites. The composition comprises primer sequences from SEQ NO:1 to SEQ NO:17 as well as block sequences from SEQ NO:18 to SEQ NO:21. The detecting method for mutations related to colorectal cancer has the advantages of high specificity, high sensitivity, low pollution, simplicity and rapidness in operation, safety and the like, detection results are better in accuracy and repeatability, the method is particularly suitable for detection of hotspot mutations of colorectal cancer driving genes from body fluids such as plasma and the like, real-time and non-invasive diagnosis, recurrence monitoring and curative effect evaluation can be performed on colorectal cancer patients, and the method has important values.

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 belongs to the technical field of in-vitro nucleic acid detection and especially relates to a relative quantitative detection method of human motor neuron gene copy numbers and a kit thereof. According to the invention, specific primers and probes to the seventh and eighth exons of SMN1 and SMN2 genes and reference gene RPP40 are respectively designed; at the same time, qualitative detection primers and probes are also designed for three hot spot mutations of Y272C, 11bp-DUP and 4bp-DEL of the SMN1 gene. The kit comprises a container containing detection primer and probe compositions, a reference container and a PCR reaction liquid container. Relative quantitative determinations to the copy numbers of the seventh and eighth exons of SMN1 (in the first and second PCR reactions) and the seventh and eighth exons of SMN2 (in the third and fourth PCR reactions) and qualitative detection to the three hot spot mutations of Y272C, 11bp-DUP and 4bp-DEL (in the fifth PCR reactions) are respectively performed by 5 independent PCR reactions. The kit is strong in specificity, high in sensitivity, convenient, fast and applicable to large-scale popularization and application.

The invention discloses a kit and a method for detecting 50 tumor-relevant hotspot mutation genes. According to the method, a capture method of 207 hotspot mutation regions of 50 relevant genes (including oncogenes and cancer suppressor genes) and a preparation method of a liquid-phase hybridization library used in capturing are researched aiming at oncology. The method particularly comprises the steps of selecting sequence information of 207 hotspot mutation regions, designing a PCR primer, preparing a biotin labeled liquid-phase hybridization library by virtue of a PCR method, hybridizing the biotin labeled liquid-phase hybridization library with a constructed target genome DNA library, and carrying out high-throughput sequencing and bioinformatics analysis on captured fragments, so as to obtain the mutation condition of nucleic acid sequence sites of a sample. By virtue of the method provided by the invention, high-quality sequencing data can be obtained, the relatively high capturing efficiency and the relatively good uniform capturing of the target sites can be realized, the disadvantages of low efficiency, poor uniformity and the like of solid phase hybridization are solved, and the sequencing cost is greatly lowered.

The invention belongs to the technical field of gene engineering, and discloses a primer combination for detecting gene mutation related to AML (acute myeloid leukemia) prognosis, a kit containing the primer combination and a method for detecting gene mutation related to AML prognosis. The primer combination covers all hotspot mutant sites of FLT3, NPM1, DNMT3A and CEBPA genes, and has the advantages of high specificity and wide covering range. The annealing temperatures of all the primers are close, so that the amplification stage can be completed at one time by one procedure, and the hotspot mutation can be subjected to combined parallel detection; and the detection efficiency is high. Besides, the forward and reverse amplification primers are adopted to directly carry out sequencing, and software is utilized to directly carry out mutation analysis, so that the method has the advantages of visual detection and low cost, thereby providing important references for therapeutic scheme determination and prognosis judgment of AML.

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 primers and a method for detecting myelosis myelodysplastic syndrome and especially detecting DOCK2 gene mutation of a patient with myelosis myelodysplastic syndrome. The primers comprise (i) primers for amplifying 6th, 22nd, 33rd, 37th, 40th and 44th exon sequences of the DOCK2 gene. An Sanger sequencing technique and sequencing primers are adopted. The primers and method can be used for quickly detecting mutation of 6th, 22nd, 33rd, 37th, 40th and 44th exons of the DOCK2 gene in the body of a patient with myelosis myelodysplastic syndrome. The primers and method have the advantage of accurate detection result, can be used for assisted diagnosis of severe combined immunodeficiency disease, and have important reference meanings for early intervention and early therapy.

The invention discloses a rapid test paper strip detection method for directly amplifying seven hot spot mutation sites (R111X, IVS4-1, Y204C, R243Q, W326X, Y356X and R413P) of a human PAH gene without extraction, and a kit. The rapid test paper strip detection method comprises: directly adding a treatment liquid to a collected sample to prepare a sample liquid; carrying out identification and amplification on SNP allele sites based on an AS-PCR method; and achieving rapid genotyping by using the interaction between amplification fragment labeled digoxin and digoxin monoclonal antibody on thesurface of nanogold magnetic microparticles and by combining with a lateral flow chromatography technology. According to the present invention, with the rapid test paper strip detection method, the DNA separation extraction process is not performed, and the surface modified magnetic gold microparticles are used as the hybrid carrier, such that the problems of high cost, time saving and labor saving due to nucleic acid purification and product treatment can be eliminated, and high sensitivity and high accuracy are achieved.

The invention discloses a primer, a probe, a locked nucleic acid probe, a kit and a detection method for detecting PDGFRA gene hotspot mutation, belonging to the technical field of biology. The primer and probe comprise at least one of the primers and probes for detecting a No.12 exon and a No.18 exon of a PDGFRA gene, the primer comprises a forward primer and a reverse primer, and the kit comprises the primers and probe. The primers and probe disclosed by the invention can be used for highly-sensitively detecting whether the PDGFRA gene has mutation, meanwhile, the sensitivity of detecting PDGFRA gene mutation can be greatly improved by adopting the locked nucleic acid probe, and whether the PDGFRA gene is mutated can be accurately detected by adopting the kit disclosed by the invention.

An ultra-sensitive, specific methodology for detecting PIK3CA mutations in biological samples of cancer patients, comprises a combination of allele-specific, asymmetric rapid PCR and melting analysis in a DNA sample from Circulating Tumor Cells, cell-free DNA in plasma / serum, or Formalin-Fixed Paraffin-Embedded tissues. Using the allele-specific primers for hotspot mutations in exons 9 and 20 (E545K and H1047R), detection can enhance amplification of mutant PIK3CA allele sequence, whereas presence of corresponding competitive blocking unlabeled probes for each exon can avoid non-specific amplification of wild-type PIK3CA sequence increasing the sensitivity and the specificity of method. The mutational detection is completed with melting curve analysis of the unlabeled probe and DNA template of the mutant PIK3CA sequence. Evaluation of PIK3CA mutational status on CTC in peripheral blood and cfDNA in plasma / serum of patients has potential for clinical applications and therapeutic interventions, since presence of PIK3CA mutations is associated with response to molecular targeted therapies.

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 embodiment of the invention provides a multiple myeloma (MM) prognosis related gene mutation detection kit and a detection method. The kit comprises multiple pairs of specific primers corresponding to the detected genes respectively; and 22 genes are selected from MM prognosis related high-mutation genes as the detection gens; according to the invention, two technologies of PCR and sequencing are comprehensively applied, rapid detection is realized for the mutation conditions of the MM prognosis related genes, specific exon sections containing hotspot mutation in the 22 detected genes are amplified through corresponding specific primers, and the mutation result is obtained through sequencing and comparison with a standard gene sequence. According to the technical scheme provided by the invention, the related genes are comprehensively selected according to the pathogenesis of MM and related signal path, and the applicability is strong; the detection specificity is high, the detection process is simple and accurate, the result is exact and clear, and the purposefulness is strong; a few experimental devices are used, and the cost is saved; and moreover, without radiolabelling, the safety is good.

The invention relates to the technical field of biology, in particular relates to medical molecular diagnosis and biotechnology, and specifically relates to a primer group and applications of the primer group. A primer combination comprises at least one group of five groups of real-time fluorescent PCR primers capable of detecting five hotspot mutation sites including MYH7-c.1987C>T, TNNI3-c.370G>C, MYH7-c2155C>T, TNNI3-c.433C>G and PRKAG2-c.298G>A, and Taqman-MGB probe sequences. The primer combination can detect a plurality of case samples at one time, also can realize the combined detection of five mutation sites on each sample, and has the features of being simple and convenient, rapid, accurate and economical, and thus a novel method is established for the clinical early molecular diagnosis and prevention of hypertrophic cardiomyopathy.

The invention belongs to the field of biotechnology and discloses a kit for detecting the hotspot mutation of KRAS gene and a detection method thereof. The kit comprises a PNA (peptide nucleic acid) reagent; and the wild type KRAS gene sequence is closed by a clamp technology so as to improve the sensitivity of a sanger sequencing process and the detection rate of the KRAS gene mutation. The detection method disclosed by the invention is simple and safe to operate and does not need many expensive reagents, thereby being economical and efficient; and meanwhile, the detection method also has the advantage of short operation time, and the whole detection process is finished within 4-6 hours.

The invention discloses a breast cancer PIK3CA hotspot mutation detection probe and primer sequence combination and a kit thereof. The detection probe and primer sequence combination comprises a primer sequence as shown in SEQ ID NO: 1-4, and a probe sequence as shown in SEQ ID NO: 5-10. The probe with the sequence is modified with a fluorophore. Based on a micro-drop digital PCR platform, primersand probes are designed for three mutation sites of the PIK3CA gene; and through experimental verification, a breast cancer PIK3CA hotspot mutation detection probe and primer sequence combination isdetermined and the combination can effectively detect the three mutation sites of the PIK3CA gene.

The invention belongs to the field of gene detection and particularly relates to a nucleic-acid mass spectrometry detection method for early screening of drive genes and susceptible genes of lung cancer. The nucleic-acid mass spectrometry detection method has the characteristics that by consideration of the difference of the lung-cancer gene spectra of the Asian populations and European-American populations, 29 lung-cancer hotspot mutations and 20 mononucleotide polymorphic sites significantly-related to suffering risk of the Asian populations are selected and combined, drive genes and susceptible genes of detection are advanced, multiple sites related to lung-cancer susceptibility and targeted-therapy drug resistance are introduced and are independent mutually without unbalanced linkage, therefore, site selection has representativeness, independence and risk-value accumulation, and can be used for evaluating the risk of individuals suffering from the lung cancer; and the detection technology is obvious in price advantage, higher in sensitivity at the aspect of lung-cancer early screening detection and larger in flux, can realize detection of multiple genes of single small samples to meet the maximum use of the small samples, and can realize translational research in clinical application.

The invention discloses a primer for detecting variation of benign and malignant related genes of thyroid nodules, a kit and a detection method. According to the primer, the kit and the detection method, variation of 15 loci of the six genes and variation of the three fusion genes can be detected simultaneously, and the BRAF gene, the KRAS gene, the HRAS gene, the NRAS gene, the TERT gene, the EIF1AX gene, RET / PTC1 fusion, RET / PTC3 fusion and PAX8 / PPARgamma fusion are involved. Hotspot mutation and fusion variation of the genes are closely related to the benign and malignant thyroid nodules. Therefore, the primer, the kit and the detection method are used for detecting a sample of a patient and can assist doctors in benign and malignant identification of the thyroid nodules which cannot be clearly diagnosed in the cytology, the accuracy of identifying the benign and malignant thyroid nodules is improved, excessive medical treatment for the patient with the thyroid nodules is reduced, and the primer, the kit and the detection method have important practical significance and high economic benefit for saving medical resources of China.

The invention relates to the technical field of biology and the field of nucleic acid detection and relates to method for detecting multi-locus low-frequency mutation of free target DNA of lung cancerplasma and a kit. The invention provides high-sensitivity and high-flux method for detecting multi-locus mutation of free DNA of lung cancer plasma once and a kit. According to the detection method,a ddPCR technique and a next generation sequencing technique are combined for detecting the mutation of lung cancer crDNA. The detection method comprises the following steps: (1) preparing a digital PCR mixed liquid including a to-be-tested sample DNA template, a primer and a PCR premixing liquid; (2) preparing a digital PCR micro-reaction drop, and carrying out PCR amplified reaction; (3) recycling and purifying a PCR product; (4) preparing and amplifying PCR premixing liquid for the second time; (5) carrying out purification and library quality control on the PCR product; and (6) carrying out computer sequencing and data analysis. According to the detection method, hotspot mutation information of 10 genes including EGFR / BRAF / KRAS / PIK3A / MET2 / ERBB2 / AKT1 / NRAS / TP53 / PTEN of lung cancer can bedetected for one time, the cost can be remarkably lowered, and the high-sensitivity and high-flue detection of mutation information of lung cancer ctDNA can be realized.

The invention discloses a hereditary deafness related gene detection chip kit, which comprises a primer and a specific probe for detecting 20 site variation on three hotspot genes of hereditary deafness, wherein 20 sites comprise 5 mutant types on GJB2, 12 mutant types on SLC26A4 and 3 mutant types on 12S rRNA (MTRNR1, belonging to a chondriogene); the sequence of the specific probe is shown as SEQ ID NO: 1 to SEQ ID NO: 40; the sequence of the specific probe is shown as SEQ ID NO: 41 to SEQ ID NO: 60. The invention also discloses a concrete detection method. The kit is designed especially forthe hotspot mutation sites of the hereditary deafness gene of people groups of southern China; the detection accurate rate of the hereditary deafness of people groups of southern China can be improved; the kit overcomes the defects that the existing kit is mainly designed for the sites of people groups of Northern China, and the detection on the people groups of southern China is liable to leak detection.

The invention discloses a primer, a kit and a detection method for detecting EGFR (Epidermal Growth Factor Receptor) gene hotspot mutation. The sequence of the primer is shown as SEQ ID NO:1-SEQ ID NO:9, the kit comprises the primer, PCR (Polymerase Chain Reaction) MIX reaction liquid, restriction endonuclease for enzyme digestion aiming at different mutations, BAS (Boric Acid Solution), enzyme digestion buffer solution, a negative control substance, a positive control substance, deionized water, bottles or tubes or packaging boxes for isolating or centrally packaging reagents. According to the primer, the kit and the detection method for detecting EGFR gene hotspot mutation, the three most common hotspot mutations of the EGFR genes can be fast detected by taking peripheral blood as a sample, the detection sensitivity is high, the time consumption is less, and the clinically needed data can be obtained in first time.

The invention discloses a primer for detecting hotspot mutation of the gene G6PD associated with glucose-6-phoshate dehydrogenase deficiency. The primer comprises primer bodies, at the sites c.1376 and c.1388, for amplifying the G6PD. By using a Sanger sequencing technology, the primer can be used for quickly detecting the mutation conditions of the sites c.1376 and c.1388 in the body of a G6PD patient. The detection result completed by using the primer is accurate, and the method has an important reference significance for rapid diagnosis of the G6PD patient.

The invention relates to the field of biotechnology, and provides a primer and a method for mass spectrometric detection of hotspot mutation of PIK3CA genes by using the primer. The method comprises the following steps of: (1) positioning hotspot mutation sites of the PIK3CA genes; (2) designing a primer for amplified reaction and a primer for extension reaction according to deoxyribonucleic acid (DNA) sequences at two ends of the mutation sites; (3) performing the amplified reaction; (4) performing treatment by using a serum amyloid protein (SAP) enzyme (alkaline phosphatase); (5) performing the extension reaction; (6) purifying a product of the extension reaction by using resin; (7) transferring the product purified by the resin to a chip; (8) putting the chip in a mass spectrograph, and running a detection procedure; and (9) reading and analyzing detection data, and determining the gene type of a target site of the PIK3CA genes. Compared with the conventional detection method, the method provided by the invention has the characteristics of high sensitivity, high accuracy, high flux and low cost.

Methods that rapidly, sensitively, and specifically detect mutations in IDH1 / 2 and the TERT promoter employ amplification of particular portions of the genes that experience frequent and exquisitely localized mutations. The ability to distinguish between sequences that differ only by one nucleotide and which may be present in very low ratios is essential for such an assay.

The invention relates to the field of medical diagnosis and particularly relates to a lung cancer multifocal diagnosis system. The system comprises a sample information processing module, a first diagnostic module, a second diagnostic module and an information output module, wherein the first diagnostic module and the second diagnostic module are used for determining whether a focus is multiple primary or metastatic according to carcinogenic driver gene and hotspot mutation of lung cancer, so that multifocal molecular determination on lung cancer can be realized.

The invention discloses an NGS-based brain tumor molecular diagnosis analysis method. The method comprises the following steps: constructing a cluster based on an Amazon cloud platform; establishing an analysis process of the molecular marker for analyzing the brain tumor on the cluster; a sequence alignment module; removing the repetitive sequence module; a gene fusion analysis module; a call variation module; a cnv analysis structure variation module; the method disclosed by the invention has the advantages of high sensitivity, high specificity, high repeatability, low experimental cost andshort analysis period, and can be used for comprehensively analyzing the hot spot mutation of the TERT promoter, the deletion states of the chromosomes 1p and 19q and other main brain tumor related molecular markers at one time. The NGS-based analytical technique of the present invention enables reliable detection of 1p and / or 19q deletions in the background tumor tissue of 70% normal cells, is more sensitive than microsatellite-based LOH analysis, and requires less DNA.

The invention provides a novel kit for detecting genetic mutation. The kit comprises a) a mutation region probe provided with a first detection marker and b) a mutation region wild type probe providedwith a second detection marker, wherein the mutation region probe can be hybridized with a mutation sequence in a hotspot mutation region of a gene; the mutation region wild type probe can be hybridized with a wild type sequence in a mutation region of the gene. A first signal of the first detection marker and a second signal of the second detection marker are obtained by using the kit, copy number and mutation frequency of the mutant gene are calculated, and whether mutation exists in the hotspot mutation region of the gene is determined.