32 results about "Dmd gene" patented technology

The invention discloses a method and a system for accurately analyzing a DMD gene structural variation breakpoint. The method includes steps of receiving of fragment sequences, initial comparison, repeat comparison, position analysis, analysis of suspected breakpoint, and in-depth correction; through the method, the type and the breakpoint position of DMD gene copy number variation can be accurately analyzed; the mean detection bias rate is less than 4 bp; the accuracy is high, and the stability is good; besides, the method and the system can cover the variation detection of an exon and an introne at the same time, solve the shortcoming that the breakpoint of the gene copy number variation cannot be accurately analyzed in the prior art, and provide technical support for realizing the rapid and parallel DMD breakpoint detection service and the study of the DMD structure gene variation molecular mechanism.

The invention relates to a method for detecting deletion and/or duplication of exons in DMD gene. The method is implemented by designing a probe using the sequence information of the known DMD gene, sequencing a DNA fragment which is obtained by capturing and enrichment, and analyzing to obtain exon deletion information of DMD gene.

The invention discloses a DMD (Duchenne muscular dystrophy) gene capture probe and application thereof in DMD gene mutation detection. A preparation method of the DMD gene capture probe disclosed herein comprises: preparing N sub-probes, connecting the sub-probes to obtain the DMD gene capture probe; N is a natural number greater than or equal to 2; the N sub-probes can cover whole sequences of DMD gene; any two adjoining sub-probes on the DMD gene meet the case where there is one or more downstream nucleotide to the corresponding upstream sub-probe, which overlap with the upstream of the corresponding downstream sub-probe; the length of each of the N sub-probes is 50-150 bp. By detecting the DMD gene with the DMD gene capture probe, it is possible to detect whether the DMD gene experiences amplification mutation deletion/repetition or not, and it is also possible to precisely locate a break point region and the size of fragments and to detect DMD gene site mutations of a sample under detection.

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 primers for detecting heterozygosity DMD gene deletion. The primers are special primers which are designed for 79 exon regions of a DMD gene and have products of 370-400 bp. Each pair of specificity primers at least have 20 bp of overlapping regions, a universal sequence and 5 continuous N basic groups are designed at the end 5' of each specificity primer, and the primers are named as labels. The invention further provides a method for detecting heterozygosity DMD gene deletion through the primers. On the basis of second-generation sequencing, by means of the method, the homozygosis and heterozygosity deletion can be detected, and therefore a carrier of the DMD genetic disease can be effectively detected, and pregnant women are guided to give birth to healthy infants through antenatal diagnosis, test tube babies and other methods.

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 relates to an amplifying system for detecting human DMD (Duchenne muscular dystrophy) gene exon copy number variation. The amplifying system comprises an amplifying composition corresponding to the detection of 10 groups of exon copy number variation, wherein the amplifying composition comprises oligonucleotide sequences as shown from SEQID No.1 to SEQID No.36; the 10 exon purpose fragments including human DMD gene exons 4,8,17,44,45,47,48,50,51,52 are amplified through a real-time multiple fluorescent quantitation PCR reaction; and according to the detection result, whether thehuman DMD gene exon is subjected to deletion mutation or repeat mutation is judged. The invention also discloses a reagent kit for detecting human DMD (Duchenne muscular dystrophy) gene exon copy number variation. The amplifying system disclosed by the invention has the beneficial effects that through the detection result, the distinguishing of DMD carriers from DMD patients and normal people canbe realized, the result is reliable, the repeatability is good, the operation is easy, the equipment requirement is low, and the reagent cost is low; and the amplifying system can cover about 90% of copy number variation in the DMD carriers or the DMD patients, so that good balance between the detection cover degree and the detection exon number is achieved.

The present disclosure provides, among other things, improved compositions and methods for treating muscular dystrophy. For example, the disclosure provides methods for treating Duchenne muscular dystrophy patients having a mutation in the DMD gene that is amenable to exon 51 skipping by administering an effective amount of eteplirsen.

The invention discloses a relative quantification method and kit for detecting the copy number of a human DMD gene through multiple real-time fluorescence PCR. The kit comprises an exon reaction solution I, an exon reaction solution II, an exon reaction solution III, a main reaction mixed solution, a positive reference substance, a negative reference substance and a blank reference substance. According to the relative quantification method and kit for detecting the copy number of the human DMD gene through the multiple real-time fluorescence PCR, the 4th, 17th and 45th exons, 8th, 50th and 52nd exons, 47th, 48th and 51st exons of the DMD gene are respectively amplified through three independent PCR reactions, and each of three reaction solutions comprises four fluorescence channels, so that the quantitative detection of the copy number of the 4th, 8th, 17th, 45th, 47th, 48th, 50th, 51st and 52nd exons of the DMD gene is realized.

The invention discloses three novel human dystrophin (DMD) gene STR gene loci and a typing method thereof, which is characterized in that the three DMD gene STR loci are respectively positioned in the 60th, the 30th and the second introns of the DMD gene and are respectively DXSDMD-in60, DXSDMD-in30 and DXSDMD-in2; and the locus fragments of the three gene STR loci have moderate sizes and are all suitable for PCR amplification. The invention selects 15 novel candidate STR sequences of the DMD gene from NCBI, uses gene typing and DNA sequence analysis and definition, studies the allele frequency distribution of the genetic structure of DMD gene STR in health populations, and obtains the DNA typing results in the health populations of the Han nationality in Xi'an city. Proven by statistical treatment, the polymorphic information contents of the three STR loci in the studied populations are all larger than 0.60, and the three STR loci can be used in population genetics and medicolegal practice and provide a theoretical basis for the gene diagnose and molecular mechanism study of Duchenne muscular dystrophy disease.

The invention provides a mini-gene splicing reporter plasmid and construction method and application thereof. A sequence of the mini-gene splicing reporter plasmid is as shown in a sequence table <400>1. Three non-continuous sequences on a DMD gene are obtained through PCR (polymerase chain reaction) and inserted to pcDNA 3.1 respectively to construct the mini-gene splicing reporter plasmid. The mini-gene splicing reporter plasmid can be used for identifying and analyzing splicing regulation elements in exons and introns and detecting whether micromutation (point mutation and microdeletion/repetition) affects splicing of the exons or not.

The invention discloses a nucleotide sequence for repairing DMD gene mutation and a system. A gene editing system can knock out a Dystrophin genomic region between the 31792310th site and the 31854834th site, or between the 31747866th site and the 31838091th site, or between the 31747866th site and the 31854834th site of X chromosome, and preferably, the gene editing system can knock out a Dystrophin genomic region between the 31815201th site and the 31846518th site, between the 31769972th site to the 31815200th site, or the 31769972th site to the 31846518th site of the X chromosome. The geneediting system can target more than 17% of DMD patients, EX51 (deficiency) 4%, EX50 (deficiency) 13%, EX45-50 (deficiency), EX48-50 (deficiency), EX50 (deficiency), EX51 (deficiency), EX52 (deficiency) and EX50 or EX51 exon area point mutation and mutation with various other forms are included, and the adaptability is higher.

The present disclosure provides, among other things, improved compositions and methods for treating muscular dystrophy. For example, the disclosure provides methods for treating Duchenne muscular dystrophy patients having a mutation in the DMD gene that is amenable to exon 53 skipping by administering an effective amount of golodirsen.

The invention discloses a DMD gene exon copy number variation detection method and application thereof and belongs to the technical field of molecular biology. 79 exons of the DMD gene are amplified in a PCR reaction solution, meanwhile, one or more than two groups of reference genes are used for monitoring the PCR reaction efficiency, PCR products are analyzed through capillary electrophoresis, calculation is carried out according to DMD female controls with the DMD genes being 2 copy numbers, relative quantitative detection is carried out on the copy numbers of the 79 exons of the DMD gene, the DMD genes with the copy numbers of 0, 1, 2 and 3 are calculated, normal exon copy number, deletion mutation and repeated mutation of the DMD gene are distinguished, and small deletion and mutation can be prompted. A result can be obtained within 4 hours from a DNA sample, the condition of the exon copy number of the DMD gene can be detected only through one-time PCR experiment, the detection technology is simple in process, and standardization is easy to achieve.

The invention discloses a duchenne muscular dystrophy (DMD) detection gene chip and kit and a use method of the kit. The gene chip comprises a solid phase carrier and an oligonucleotide probe fixed on the solid phase carrier, and the oligonucleotide probe is indicated in a nucleotide sequence of SEQ ID No. 1-345; the gene chip is contained in the kit; the use method of the kit includes the steps of a), preparing sample DNA fragments; b), fluorescently marking the DNA fragments; c), purifying gDNA markers; d), allowing the gDNA markers to be interbred with the gene chip; e), cleaning the interbred gene chip and scanning chip interbreeding signals to acquire results. The gene chip and the kit are low in cost, simple in detection operations, high in specificity and good in stability, the process from sample extraction to acquisition of the scanning results can be completed within two days, and the gene chip and the kit are suitable for detection for one or multiple exon deletion or repeat mutations in clinical patient DMA gene and antenatal diagnosis.

The invention discloses application of a histone deacetylase inhibitor in preparation of a medicine for treating heart diseases, and belongs to the technical field of medical application of compounds. A mouse model with CDK13 knocked out of myocardium is constructed, and it is confirmed that deficiency of CDK13 in myocardium can cause cardiac dysfunctions such as dilated cardiomyopathy and heart failure; a molecular mechanism that CDK13 causes cardiomyopathy by regulating down-regulation of DMD gene expression is found. A histone deacetylase inhibitor No.1217 (the molecular formula of the histone deacetylase inhibitor No.1217 is C24H27N3O4) is used for performing drug treatment on a myocardial knockout CDK13 mouse, and the result shows that after continuous administration is performed for one month, the cardiac function, the cardiac fibrosis pathological process and the dystrophy protein level of the myocardial knockout CDK13 mouse are partially reversed. The invention finds an effective molecule and a treatment strategy for treating cardiomyopathy by targeting CDK13, and has important significance for treating heart disease.

The invention discloses a test kit of preimplantation embryonic genetics diagnosis and prenatal diagnosis for DMD, which comprises a DMD gene linked SNP site capture probe group, the DMD gene linked SNP site refers to 199 or more SNPs sites which are within 2M of the upstream and downstream of the DMD gene and have high mutation frequency in people. The probe group in the test kit disclosed by theinvention can be used for rapidly and accurately capturing the DMD gene and the DMD gene linked SNP at the same time, and the detection specificity and accuracy of the DMD preimplantation embryonic genetic diagnosis can be improved.

The invention discloses application of Ozanimod in preparation of a medicine for treating muscular dystrophy, and belongs to the field of medicine. An internationally recognized DMD gene knockout mouse, namely an mdx mouse, is used as a muscular dystrophy model, and the treatment effect of ozanimod on myasthenia and amyotrophy caused by muscular dystrophy is investigated on the mdx mouse. The influence of ozanimod on myasthenia and amyotrophy caused by muscular dystrophy is systematically studied from the aspects of pharmacodynamics, behavioristics, molecular biology and the like, and the result shows that ozanimod can reverse the deficiency of muscle fibers and enhance the strength and function of skeletal muscles.