150 results about "Pathogenic mutation" patented technology

The invention discloses a DNA (Deoxyribonucleic Acid) library for detecting cholestatic jaundice pathogenic genes by virtue of a high-throughput sequencing technology and application thereof and in particular relates to a super-multiplet PCR primer which can cover the exon and adjacent areas of the genes according to 60 cholestatic jaundice pathogenic genes. The genome DNA of the sample is subjected to super-multiplet PCR amplification, the amplification product is sequenced by utilizing the high-throughput sequencing technology, the pathogenic mutation is searched, the genetic etiology of the cholestatic jaundice is clear, and a theoretical basis of genetics and molecular biology is provided for clinical diagnosis. The DNA library has the characteristics of accuracy, flexibility, rapidness and low cost. Moreover, according to the detection area of 60 genes, 15 common cholestatic genetic defects can be detected, and the DNA library has significance and clinical values on differential diagnosis of the cholestatic jaundice.

The invention discloses a targeting-based new generation sequencing deafness gene detection set and kit, and a detection method. The deafness gene detection set comprises 258 genes, 81 non CDS regions and a whole mitochondrial group. The detection method comprises the steps: designing primers for a part of or all of deafness disease genes and loci; with a to-be-tested sample DNA as a template, carrying out PCR amplification with the primers, and constructing a deafness detection gene library based on the amplified product; and according to the deafness detection gene library, establishing a sequencing template, carrying out high-throughput sequencing, and analyzing sequencing data information. The invention also discloses the related kit. Not only can conventional known mutations be detected out, but also new mutation types also can be detected out; in addition, sequencing and analysis of a large number of objective regions also can be completed within a short period of time, and positions, possible to generate pathogenic mutation, of all exons and regulatory regions of the hundreds of genes associated with deafness are subjected to related sequencing and analyzing.

The invention provides a whole-exome sequencing data analysis method. The method comprises the following steps of 1) quality control of sequencing data; 2) genome mapping of the sequencing data; 3) seeking of high-confidence genome mutation by the sequencing data; and 4) annotation of mutation sites. According to the method, the analysis of large-scale data is finished through simple parameter submitting, wherein the analysis of the large-scale data comprises quality detection of original data, data denoising and genome mapping of sequencing read; an upstream part takes over original sequencing data of a lower machine; the analysis of the sequencing data is finished through a parameter automated submitting and analysis module; and candidate pathogenic mutation sites and related genes are output, thereby providing a basis for later experimental verification.

The invention discloses a DNA library for detection of hereditary cardiomyopathy causing gene mutation through the targeting high-flux semiconductor sequencing technology and application thereof. Specifically, a primer pool is designed according to 80 hereditary cardiomyopathy causing genes, super-multiple PCR amplification is performed on sample genome DNA, and an amplified product is sequenced through the high-flux semiconductor sequencing technology to find pathogenic mutation so as to provide genetic and molecular biological theoretical bases for clinical diagnosis. The DNA library has the advantages of being accurate, quick, flexible and low in cost; through 80 gene detection areas related to the DNA library, five kinds of common hereditary cardiomyopathy including hypertrophic cardiomyopathy, dilated cardiomyopathy, arrhythmogenic right ventricular cardiomyopathy, restrictive cardiomyopathy and left ventricular myocardial densification incomplete cardiomyopathy can be detected, and the DNA library has an important meaning and clinical value for diagnosis and differential diagnosis of the hereditary cardiomyopathy.

The invention provides an exome potential pathogenic mutation detection method based on a family line. The detection method comprises the following steps: 1) reading a result file of an exome sequencing data processing flow, and conducting function filtering; 2) reading the file obtained in the last step, extracting mutations in all samples, calculating a union set, and then combining all samples, so that a matrix is constituted; 3) extracting mutation information in the matrix obtained in the last step, enumerating and assessing pathogenicity of single mutation and pathogenicity of combined dual-site mutation, so that a potential pathogenic mutation list is obtained; and 4) in accordance with the list obtained in the last step, calculating the appearance situations of sites in various samples and target genes. According to the method disclosed by the invention, data integration and basic filtration are completed by taking an output result of the common exome sequencing processing flow as an input condition; by virtue of a special mutation screening algorithm, a candidate set of the potential pathogenic mutations is provided; and the method focuses on solving a problem on potential pathogenic mutation mining of sequencing data with high heterogeneity, high mutation rate and high noise.

The invention discloses a probe and a kit for detecting pseudo-hypertrophic muscular dystrophy. The probe for detecting pseudo-hypertrophic muscular dystrophy comprises nucleic acid sequences shown by SEQ NO:1-SEQ ID NO:546, and the kit for detecting pseudo-hypertrophic muscular dystrophy comprises nucleic acid sequences shown by SEQ ID NO:1-SEQ ID NO:546. The probe and the kit for detecting pseudo-hypertrophic muscular dystrophy can be used for detecting complete mutation information of DMD genes, including exon deletion/repetition and point mutation, have the advantages of completeness, rapidness, accuracy and appropriate price, and can be used for solving the problems of DMD molecular diagnosis; the kit disclosed by the invention is suitable for detection of various tissue samples of muscle, blood, amniotic fluid and the like, can detect pathogenic mutation of exons and introns as well as known mutations and unreported pathogenic mutations, and is also used for detecting whether patient genotypes are corrected or not after stem cell therapy.

The invention discloses a multi-factor model for predicting a breast cancer onset risk and an establishing method of the multi-factor model. The establishing method of the multi-factor model comprises screening gene detection, wherein the screening comprises the following steps: (1) collecting and treating a peripheral blood sample; (2) establishing library DNA of the sample, and performing computer sequencing on a Hiseq sequencing platform; (3) performing biological information analysis on computer data, and finding pathogenic mutation; (4) sequencing mutation sites by using a golden standard Sanger sequencing technique, thereby obtaining accurate gene mutation information. By adopting the multi-factor model which is disclosed by the invention and is used for predicting the breast cancer onset risk, a part of people can be screened and diagnosed at an early stage, active interference measures can be taken, and the multi-factor model is particularly applicable to prediction on high-risk people suffering from familial breast cancer.

The invention discloses a DNA library for detecting pathogenic genes of multiple genetic vascular diseases through a targeted high-throughput semiconductor sequencing technology and application thereof. The operating method specifically comprises the following steps: designing a primer pool according to 151 pathogenic genes of genetic vascular diseases, performing super-multiple PCR amplification on genomic DNA of samples, sequencing the amplification products by utilizing the high-throughput semiconductor sequencing technology, searching pathogenic mutations, and providing genetic and molecular biological theoretical basis for clinical diagnosis. The DNA library disclosed by the invention has the characteristics of accuracy, rapidness, flexibility and low cost, the 151 gene detection areas involved in the invention are capable of detecting multiple genetic vascular diseases such as aortic dissection, arterial aneurysm, hemorrhagic apoplexy and the like, and the DNA library has great significance and clinical values on diagnosis and differential diagnosis of the genetic vascular diseases.

The invention belongs to the field of genetic engineering and tumor medical science and discloses a BRCA2 gene g.32912799T>C mutation and application of the mutation in auxiliary breast cancer diagnosis. Compared with a normal BRCA2 gene sequence, the BRCA2 gene mutation has a g.32912799T>C locus mutation. The BRCA2 gene g.32912799T>C mutation provides a new pathogenic mutation locus of breast cancer and can be used for diagnosing breast cancer at the early stage.

The invention relates to a detection system of an SMA (spinal muscular atrophy) related gene and a detection kit and belongs to the clinical molecular diagnosis technology in the biomedical field. Thedetection system can simultaneously amplify and detect 2 SMN gene pathogenic mutation sites and 3 contrast sites in one multiple compound PCR amplification system. The detection system has high sensitivity and high resolution and can clearly and effectively distinguish 1 bp difference in the general 100-500 bp detection range; the detection speed is high, required operation is few, automatic massdetection can be realized, the operation intensity and detection cost can be greatly reduced, and results can be obtained within 4 h. Detection samples can be amplified by blood, blood cards and other detection materials, a DNA extraction step is omitted, the operation is more convenient, and the system is suitable for mass operation. As supplement of detection of SMN1 seventh exons, the detection has important application values for SMA study and carrier screening.

The invention relates to a gene detection probe, a primer and a kit for spinal muscular atrophy, wherein the gene detection probe for spinal muscular atrophy comprises a probe SMN1-7 for detecting a SMN1 gene, and the nucleotide sequence is shown in SEQ ID No. 1; a probe SMN2-7 for detecting a SMN2 gene, and the nucleotide sequence is shown in SEQ ID No. 2. According to the invention, a multiple fluorescence quantitative detection system with high specificity and low cost is established. High frequency pathogenic mutation detection of the SMN1 and SMN2 genes in one tube is realized. In addition, a multiple scorpion tail primer amplification system is established, so that the problem of the amplification consistency of the multiple fluorescence quantitative systems is solved, and the quantitative accuracy is improved.

The invention discloses a pathogenic mutation of osteogenesis imperfecta and a detection reagent of the pathogenic mutation. According to a novel mutant COL1A1 gene, the mutated COL1A1 gene is a single point mutation c.1822G>A (chr17:48270211), a heterozygous mutation is pathogenic and in a mode of dominant heredity, the amino acid change is p.Gly608Ser, dyssynthesis of I-type collagen in connective tissue is caused by locus mutation of the p.Gly608Ser, and a lesion is formed. A kit for detection of osteogenesis imperfecta includes a reagent for detection of the 1822bpth locus of a COL1A1 geneCDS or a reagent for detection of the 608th amino acid locus of a COL1A1 protein. The pathogenic mutation (c.1822G>A on the COL1A1 gene) of the osteogenesis imperfecta disease is obtained, and the osteogenesis imperfecta disease can be diagnosed by detecting the mutation.

The invention discloses a genetic diagnosis kit of Alzheimer's disease. The kit can detect, research and report 10 virulence genes related to Alzheimer's disease, including APP, APOE, PSEN1, PSEN2, GSK3beta, DYRK1A, Tomm40, CLU, PICALM and TAU. A detecting method comprises the following steps: designing and synthesizing specific primers of all genes, collecting the sample of a individual to be detected, adopting the RT-PCR technology to carry out sequencing analysis on obtained specificity DNA products of all genes, comparing the sample sequence with the normal gene sequence, and analyzing whether pathogenic mutations exist to evaluate the morbidity risk of the individual. The genetic diagnosis kit of Alzheimer's disease can detect different pathogenic mutation sites of CDS at a time and has the advantages of high simpleness, convenience, rapidity and sensitivity, reliable preparation and excellent specificity.

The invention relates to a detection primer, a detection kit and a detection method of Leber's hereditary optic neuropathy (LHON) mitochondrial DNA gene mutations. The invention, on the basis of Sanger sequencing principle, has the characteristics of being highly sensitive, stable and accurate; four most common pathogenic mutations of LHON mtDNA can be simultaneously detected, so that the problems of an existing LHON mitochondrial DNA mutation detection method, which is low in sensitivity, not strong enough in specificity, capable of causing pollution easily, high in expense and the like, can be solved; therefore, an accurate gene diagnosis method is provided for LHON; and the invention has important implications for genetic counseling and gene therapy of the disease (the LHON).

The invention discloses a multiplex PCR (Polymerase Chain Reaction) trapping primer group for a DMD (Duchenne Muscular Dystrophy) gene, a mutation detection kit for the DMD gene and a mutation detection method for the DMD gene. The inventors provide the multiplex PCR trapping primer aiming at the sequences of 79 exons and bilateral 50bp introns and 23 known intron pathogenic mutations of the DMD gene; moreover, the mutation detection method of the DMD gene is provided based on the primer group; the mutation detection method is high in detection sensitivity, good in accuracy, highly consistentin amplification efficiency and low in requirement on quality of a DNA (Deoxyribonucleic Acid) sample, and is used for solving the problem that the complete detection on the mutation information, particularly on loss of heterozygosity/repetition, of the DMD gene is difficultly carried out in one time by adopting a multiplex PCR trapping technique in the prior art.

The invention discloses an amplification primer for detecting PAH (phenylalanine hydroxylase) gene mutation, a kit and a detection method thereof. A primer group for detecting PAH gene mutation through PCR (polymerase chain reaction) specific amplification comprises 9 pairs which are respectively primers used for amplifying PAH gene exon 1 upstream lateral sequences (positioned at the gene 5' end)to an introne 2 (a forward primer is shown as SEQ ID NO:1, and a reverse primer is shown as SEQ ID NO:2), primers are used for amplifying PAH gene introne 2 upstream (5' end) to introne 2 downstream(3' end) (a forward primer is shown as SEQ ID NO:3, and a reverse primer is shown as SEQ ID NO:4), and the like. The PAH gene whole coverage is realized; the detected sequence length reaches 88171bp;the intron variation and structure variation detection can be facilitated; the detection rate of pathogenic mutation can be improved; the operation is simpler; the cost is low; in addition, each amplification fragment and the adjacent fragment have the overlapping region; the primers can be used for fragment splicing and haplotype construction.

The invention belongs to the field of gene treatment, and specifically relates to a method adopting a base editing system to repair beta thalassemia caused by A>G pathogenic mutation. The provided base editing system specifically repairs the above mutational method, mutational gRNA above specificity targeting and base editing protein. HBB: c.-79A>G and HBB: c.-78 A>G of human can be accurately repaired by utilizing the method. The method is high in efficiency and good in specificity. The gene editing system can be introduced into a somatic cell or individual of human, accurate repair can be performed on the A>G pathogenic mutation, thereby beta thalassemia disease can be cured, and the technology has wide application prospect in the field of gene treatment.

The invention discloses a construction method of an Adgrv1 gene Y6236fsX1 mutant animal model based on CRISPR/Cas 9 technology. The method comprises the following steps: (1) designing and choosing theGuide RNA (ribonucleic acid) and repair template DNA (deoxyribonucleic acid) of proper target mouse Adgrv1 genes; (2) synthesizing Cas9 mRNA, the Guide RNA and the repair template DNA in vitro; (3) enabling the Guide RNA, the Cas9 mRNA and the repair template DNA to be jointly prepared into an injection sample in a mole ratio so as to be injected into a fertilized ovum; (4) identifying obtained F0-generation mice; and (5) enabling positive F0-generation mice to perform passage and obtaining and identifying F1-generation mice. The method can be used for successfully obtaining an Adgrv1 gene Y6236fsX1 mutant mouse model corresponding to Y6244fsX1 pathogenic mutations in human, thereby laying a foundation for researching relevant factor and diagnostic treatment of the Y6244fsX1 pathogenic mutations in human.

The invention provides a method and kit for rapidly detecting abnormal GGC repeat amplification of NOTCH2NLC genes. The components of the kit include a reaction solution, a reinforcing agent, a specific primer, dNTPs and DNA polymerase. Through a high GC content PCR system, GGC can be repeatedly amplified through the DNA polymerase of high thermal stability and high GC content templates, so that the number of GGC repeat of to-be-detected samples can be calculated through fluorescence scanning, and whether amplification occurs can be judged as well. The kit can detect the repeat number of NOTCH2NLC gene GGC through the PCR system 1, determine whether the repeat number is in a normal range or a mutation range, and screen whether subjects carry GGC pathogenic mutation; and whether amplification exists can be judged through the trapezoidal peak of a PCR system 2, so that missed detection can be avoided. The method can be applied to neuronal intranuclear inclusion diseases and the gene detection in some dementia patients. The kit has the characteristics of being fast, accurate and high insensitivity, and can prevent missed detection.

The invention discloses a deafness-related gene capture kit and an application thereof. The invention provides a capture probe. The capture probe is as shown in 1) or 2) which are described in the specification, wherein 1) the capture probe is composed of probes as shown in sequences 1 to 113 in a sequence table; 2) the capture probe is composed of derivatives of probes in 1); and the derivative of each probe is a probe which is obtained by substituting and/or deleting and/or adding one or more nucleotides to each probe in 1) and has the same function. According to the deafness-related gene capture kit provided by the invention, target region sequencing analysis is carried out on a whole exon group or a deafness-related genome (respectively accounting for 1% and 0.01% of the whole genome)of a hereditary deafness patient; and most pathogenic mutation information of the disease is captured.

The invention belongs to the fields of gene engineering and tumor medical science, and discloses a mutant gene for breast cancer auxiliary diagnosis and application of the mutant gene. The mutant gene has one or more mutant sites: a mutant site A which has g.41215910delA, g.41244291delT, g.41223130A>T and g.41256139T>A, and other mutation sites which have g.32912699A>G, g.32912799T>C, g.32929399dupT, g.32911757C>T, g.32911659-32911662delAAAA and g.32910963T>G compared with a normal BRCA2 gene sequence. The invention provides a breast cancer pathogenic mutation site combination, and the mutation site combination can be used for early diagnosis of the breast cancer.

The invention relates to a DNA library for detecting and diagnosing pathogenic genes of skeletal development disorders through a targeted high-throughput sequencing technology and application thereof. The library comprises 507 pathogenic genes of skeletal development disorders. According to the invention, 507 pathogenic genes of skeletal development disorders are preferably selected, a probe pool is designed, a target region library for 507 pathogenic genes of skeletal development disorders is established, and the library utilizes the high-throughput sequencing technology for sequencing to find pathogenic mutations, thereby providing genetic and molecular biological bases for clinical diagnosis. The DNA library provided by the invention has the characteristics of accuracy, rapidness, flexibility and low cost. The 507 genes involved in the invention include pathogenic genes of genetic diseases with skeletal development disorders as clinical manifestations, such as collagen dysplasia, metaphysic dysplasia, osteogenesis imperfecta and bone density reduction, mucopolysaccharide storage disease, cartilage dysplasia and the like, and have important significance and clinical value for diagnosis, differential diagnosis and accurate treatment of skeletal development disorders.