78 results about "Hereditary deafness" patented technology

The present invention provides a chip, a kit containing the chip, uses of the kit in hereditary deafness detection, a deafness related gene detection method, and a deafness related gene detection apparatus. According to the present invention, with the chip, the kit, the method and / or the apparatus, the deafness related gene region can be completely captured in one time, and the variation condition of the deafness related gene can be detected.

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 relates to a hereditable deafness susceptibility gene screen test kit. The kit uses mutation from C to T of 1,494 locus of a 12s rRNA gene, mutation from A to G of 1,555 locus of the 12srRNA gene, mutation from A to G of IVS7(-2) locus of an SLC26A4 gene and C(+ / -) of 235 locus of a GJB2 gene as detecting objects, designs and optimizes a set of specific primers respectively againsteach locus to be tested according to the PCR technical principle of a tetra-primer amplification refractory mutation system, amplifies the whole set of the primers in the same reaction tube, and performs primary multiple PCR amplification and primary gel electrophoresis on the four reaction tubes to obtain gene types of four loci simultaneously.

The invention discloses a primer system which is used for simultaneously detecting 10 gene SNPs (single nucleotide polymorphisms) related to hereditary hearing loss. Based on a product prepared by using the primer system, the 10 gene SNPs related to the related to hereditary hearing loss can be simultaneously detected. The product can be used for detecting the gene types of 10 gene SNPs of a testee, and the detection results can be used as a reference to clinical diagnosis and also can be used in the fields, such as epidemiological investigation, prenatal screening and neonatal screening. According to the primer system, 10 gene SNPs on different genes can be simultaneously detected in one reaction system; and therefore compared with technologies, such as sequencing, real-time fluorogenic quantitative PCR (polymerase chain reaction), the primer system is lower in cost and simple and convenient to operate, and the accuracy and sensitivity are improved.

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 brings forward a kit for detecting hereditary hearing loss. The kit provided by the invention comprises a primer combination and a probe combination of nine hereditary hearing loss polymorphic sites on GJB2(Cx26) gene, SLC26A4(PDS) gene and 12S rRNA(MTRNR1) gene, and can be used to accurately determine the wild type, homozygous mutation or heterzygosity of related sites and realize molecular diagnosis of hearing loss. With the application of the kit provided by the invention, detection can be carried out by cheap equipment or macroscopic observation. The primer combination, reagent panel and kit provided by the invention can be used for the screening of hereditary hearing loss to realize early detection, early prevention and prenatal guidance. The invention is of great commercial value and social significance.

The present invention discloses a primer system for concurrently detecting 10 gene polymorphism sites related to genetic deafness. Based on the product prepared by the primer system, 10 gene polymorphism sites related to genetic deafness can be concurrently detected. Use of the product, genotypes of 10 gene polymorphism sites of a subject are detected, and the detection results can be used for assisted clinical diagnosis, and can further be used for epidemiological investigation, and can also be used for prenatal screening, neonate screening and other fields. With the present invention, 10 gene polymorphism sites on different genes can be concurrently detected in a reaction system, such that advantages of low cost, convenient operation, accuracy improvement and sensitivity improvement are provided compared with sequencing, real-time fluorescence quantitative PCR and other technologies.

The invention relates to the field of gene detection, in particular to a nucleic acid membrane strip and a kit for hereditary hearing loss gene detection. The kit comprises a PCR reaction liquid, wherein the PCR reaction liquid comprises a PCR reaction liquid A, a PCR reaction liquid B, a PCR reaction liquid C, a PCR reaction liquid D and a PCR reaction liquid E and also comprises corresponding primers and probes. According to the kit, the specific amplification LATE-PCR special primers are designed by use of the sequence of various mutation sites, reported by in literature, and the locked nucleic acid modified ZNATM probes are adopted, so that the least tubes of the reaction liquid can detect the most sites. The PCR reaction liquids A, B, C, D and E in the kit can be tested on the same fluorescent PCR instrument by use of the same amplification program, and the clinical requirement for quick and convenient hearing loss detection is satisfied.

The invention relates to a kit for building a library for detecting hereditary deafness genes and its application. The kit includes an amplification primer set for amplifying hereditary deafness genes; Primers designed for the polymorphism of the gene and the mutation site of the 12s rRNA gene. The present invention adopts multiplex PCR technology, which can detect more than 20 mutation sites of deafness genes in a single reaction; in conjunction with the "double label" system, the DNA amplification product of each sample has two sets of independent label sequences, so The amplification products of all samples can be mixed and sequenced simultaneously to achieve high-throughput detection, thereby greatly reducing the detection cost of a single sample.

The invention a mutation detection kit used for performing qualitative detection on 20 gene loci related to hearing loss in a genome DNA. The kit comprises a DNA diluent, a hybridization reaction liquid, a hearing loss probe mixed liquid A, a hearing loss probe mixed liquid B, a connecting reaction liquid A, ligase, a connecting reaction liquid B, polymerase, a PCR amplification primer mixed liquid, positive control, and negative control. The kit provided by the invention performs gene mutation detection mainly by combination of a multiplex ligation-dependent probe amplification (MLPA) technology and capillary electrophoresis, so that multiple mutation loci can be detected at the same time; wild type, homozygous mutation type and heterozygosis mutation type of 20 loci can be accurately determined by two tubes concurrently; and meanwhile, a primer probe is also designed, so that quite high accuracy and specificity are achieved.

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

The invention relates to a method for testing gap linking protein 26 gene GJB2 mutation related to hereditary hearing impairment, which completely enlarges GJB2 gene base boot sector, exon 1, exon 2 and shear zone by polymerase chain reaction; and then, DNA sequence is measured to detect whether GJB2 genetic mutation exists; the method of the invention can completely cover all the genetic mutation of the GJB2 gene base boot sector, the exon 1, the exon 2 and the shear zone, so that detectable rate of the GJB2 genetic mutation and diagnosis rate of hereditary hearing impairment related to the GJB2 gene are improved; compared with that sequence measurement is separately carried out on a GJB2 code area, the method is more comprehensive and reliable, so as to be beneficial to the diagnosis of hereditary hearing impairment.

The invention relates to a multiplex ligation-dependent probe amplification kit for detecting hereditary deafness susceptibility gene mutation. The multiplex ligation-dependent probe amplification kit comprises a detection probe for detecting mutation site specificity, a universal primer, a positive DNA template, a DNA ligase, a connection buffer solution, a hybridization solution and a PCR reaction reagent, wherein the detection probe is composed of a pair of short probe and long probe and corresponds to one mutation site. The method is high in sensitivity and is accurate in result, and compared with the traditional detection technology, the kit can be used for completing high-throughput screening of multiple key hereditary deafness gene mutation diagnosis at one time, and has the advantages of high economical efficiency, high efficiency and high accuracy.

The invention relates to a gene chip for non-invasive prenatal diagnosis of high-risk hereditary hearing loss and a preparation method in the technical field of gene diagnosis. The gene chip comprises a substrate and probes fixed on the substrate, wherein the probes are nucleotide sequences shown as SEQ ID NO.1-24. Usage of the gene chip includes the steps of sample preparation, multiplex PCR (polymerase chain reaction) amplification, ligase detection reaction, chip hybridization, chip scanning and result obtaining. Beside, the invention further relates to a reagent kit comprising the gene chip. The gene chip for diagnosis is simple in operation steps, only requires the multiplex PCR reaction, the ligase detection reaction and hybrid scanning once, is high in detection specificity and good in stability so as to be capable of correctly distinguishing homozygotes from heterozygotes of each locus and high in repeatability of repeated experiments, is short in detection time and low in cost as the substrate can be universally used for other chips by means of universal chip technology.

The invention discloses a probe, a primer, a detection reagent kit and a detection method for detecting hereditary hearing impairment on the basis of a floating-bead array system. The probe used for mutation of detecting hereditary hearing impairment genes is coupled and mixed with microbeads of different codes to acquire a floating-bead array; the specific primer is used for performing multiple PCR (polymerase chain reaction) amplification, products of the floating-bead array is subjected to hybridization with the floating-bead array, streptavidin R-phycoerythrin is used for color developing, and detecting results are read through a floating-bead array reading instrument. By the arrangement of the probe and the primer for detecting hereditary hearing impairment on the basis of the floating-bead array system, 20 mutation sites of the four major hereditary hearing impairment genes can be detected simultaneously, detection time can be greatly shortened, and detecting efficiency and accuracy can be improved.

The invention belongs to the field of gene detection and especially relates to multiple PCR specific primers, kit and method for detecting a hereditary hearing loss gene, based on a high throughput sequencing technique. The primers include 52 pairs of specific primers, the nucleotide sequences of which are shown as SEQ ID NO:1 to SEQ ID NO:104. The kit further comprises the specific primers, a universal primer, an enzyme mixture, a quality control substance and nuclease-less water. The invention also discloses a detection method for capturing and amplifying the hereditary hearing loss gene through once PCR amplified reaction by utilizing the primers or kit. According to the scheme provided by the invention, the detection efficiency is effectively increased, the accuracy is increased, the cost is lowered and the operation step is simplified.

The invention discloses an HRM method and kit for clinically detecting deafness-related gene mutation. The HRM method comprises the steps that PCR amplification and HRM scan analysis are conducted by extracting DNA of a sample to be detected and applying primers shown in SEQ ID NO.1-30; the deafness-related gene mutation type is judged according to a scan analysis result, a sample free of mutation peaks in HRM scan typing is negative, and a sample with the mutation peaks in HRM scan typing is positive. The kit comprises the primers, quality control products, a PCR reagent, fluorescent dyes and the like; the kit has the advantages that all the primers in the kit can conduct a PCR reaction at the same temperature, the detection sensitivity and the specificity are high, a small reaction system is achieved, the detection sample source is rich, reacting is easy, convenient and rapid, the reagent cost is low, and high throughput is achieved. The HRM method and kit are suitable for clinical neonatal hereditary deafness gene screening and pre-pregnancy deafness gene screening for good prenatal and postnatal care and beneficial for guiding good prenatal and postnatal care and clinical personalized medication.

The invention relates to the field of gene detection and particularly relates to a nucleic acid membrane strip and a kit for detecting a gene of hereditary hearing loss. The nucleic acid membrane strip comprises a substrate and specific oligonucleotide probes which are fixed on the substrate, wherein the sequences of the oligonucleotide probes are shown in SEQ ID NO: 1-42. According to another technical scheme, the invention provides a kit for detecting the gene of hereditary hearing loss. The kit comprises the nucleic acid membrane strip detecting the gene of hereditary hearing loss, a PCR reaction liquid I and a PCR reaction liquid II. The nucleic acid membrane strip and kit provided by the invention have the beneficial effects that the nucleic acid membrane strip and kit comprise eight pairs of primers capable of being efficiently and specifically amplified and 42 probes capable of being specifically hybridized. The detection sites are relatively complete, so that 21 common hearing loss sites can be simultaneously detected by using two tubes of the reaction liquid and one membrane strip.

The invention discloses a hereditary hearing loss susceptible gene 20 site typing detection kit. The kit comprises PCR amplification primers and single-base extension primers of 1494 C>T and 1555 A>G sites of a mitochondrial gene, 35dleG, 299-300delAT, 235delC, 176-191del19 and 167delT sites of a GJB2 gene, 547G>A and 538C>T sites of a GJB3 gene, and 281C>T, 589G>A, IVS7-2A>G, 1174A>T, 1226G>A, IVS15+5G>A, 1975G>C, 2027T>A, 2162C>T and 2168A>G sites of a SLC26A4 gene. The primers have sequences shown in the formulas of SEQ ID No: 1 to SEQ ID NO: 40. The kit is divided into two groups and is used for amplification and detection of 20 sites so that the accuracy of detection is guaranteed. The kit realizes high efficiency, low cost and high accuracy detection of a hereditary hearing loss susceptible gene type.

The invention discloses a Cpf1 kit for rapid detection of hereditary deafness pathogenic gene SLC26A4 mutation and a detection method thereof. The Cpf1 kit comprises a Cpf1 detection system. The Cpf1detection system comprises: a specific crRNA for SLC26A4, a Cpf1 protein and a single-stranded DNA reporting system. The specific crRNA is any one or more designed and synthesized for mutation sites.The single-stranded DNA reporting system comprises an ssDNA FQ reporter for fluorescence detection by an enzyme-labeled instrument and / or an ssDNA DB reporter for immune colloidal gold strip detection. By detecting SLC26A4 gene mutation sites with the Cpf1 for the first time, the kit of the invention has the advantages of high sensitivity, strong specificity, low time consumption, no dependence onlarge-scale experimental equipment and the like.

The invention discloses a genetic deafness associated gene detection kit and a specific primer set. The invention designs a specific primer set for 33 mutation sites of genetic deafness-related genes, and a detection kit for genes associated with genetic deafness was prepared, the kit and the specific primer set of the present invention can be used quickly, Accurate, one-time detection of 33 genetic mutations associated with genetic deafness, and the use of specific primers for the construction of a library of multiple PCR homogeneity and stability, the amount of required initial template DNA, the library obtained from the utilization of sequencing data and detection accuracy is high, applicable to a variety of sample types.

The invention provides a detection kit for c.403>T mutation of the CDH23 gene. Causes and types of hereditary hearing loss and / or Usher ID syndrome are diagnosed through detecting whether patients have the mutation gene or not. The mutation gene and a detection method mentioned above are beneficial for clinical screening of CDH23 mutation for patients with hereditary hearing loss, providing bases for diagnosis and treatment of patients with hereditary hearing loss and Usher ID syndrome.

The invention provides a reagent kit for simultaneously detecting multiple deafness genes on a single cell level. The invention also provides a method for simultaneously detecting various deafness gene sites on a single cell level. By adopting a single-cell whole genome amplification technology, a sample type with little content of DNA such as a single-cell sample is effectively amplified to whole-genome DNA with total amount of micro-gram by virtue of MDA (multiple displacement amplification). The obtained DNA is amplified by virtue of PCR of a specific primer, Sanger sequencing is carried out for an amplified product on a sequencing instrument, and various genetic deafness genes can be effectively detected.

The invention discloses a reconstructed oocyte of a deaf model pig and a construction method of the reconstructed oocyte, and also discloses the deaf model pig and a construction method and application of the deaf model pig. According to the reconstructed oocyte of the deaf model pig, the construction method of the reconstructed oocyte and the construction method of the deaf model pig, by knockingout a pig OSBPL2 gene, the reconstructed oocyte of a deaf mini-pig is obtained, and in combination with a CRISPR / Cas9 gene editing technology, a somatic cell nuclear transfer technology and an embryotransfer technology, high efficiency and feasibility of the construction method for the genetically engineered deaf model pig are proven. Through the construction method of the deaf model pig, genotypes and deafness phenotypes of human OSBPL2 gene mutations can be precisely duplicated, the pig of which an OSBPL2 gene is knocked out has typical characteristics of human autosomal dominant hereditary hearing loss (DFNA67), therefore, a reliable big animal model can be provided for disease research of the human hereditary hearing loss, and the deaf model pig can be applied to inner-ear gene repairing, inner-ear hair cell regeneration, hearing reconstruction and the like.

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 provides a primer group, a method and application of a hereditary hearing loss screening based on a MassArray nucleic acid mass spectrum platform. The primer group for detecting SNP sites related to hereditary hearing loss and a detection system thereof are designed on the basis of the MassArray nucleic acid mass spectrum platform, so that mutation types such as point mutation, deletion mutation and mitochondrial point mutation and the like related to hereditary hearing loss diseases can be accurately typed, and rapid and effective detection of hereditary hearing loss diseases isrealized.

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 field of biomedicine and gene engineering. Probe combination is disclosed by the invention and can be used for identifying exon sequences of pathogenic genes GJB2, GJB3 and SLC26A4 aiming at hereditary hearing loss or SNP sites in upstream or downstream 2Mb of the genes. The probe combination is suitable for crowd diagnosis and screening, and comprises non-invasive prenatal detection or embryo pre-implantation detection. The invention also discloses a corresponding detection method, a detection kit and an application thereof. According to the invention, hereditaryhearing loss detection can be carried out, detection of all exons and splicing areas of pathogenic genes is realized, the coverage area is more comprehensive, family linkage analysis can be realized by combining genes adjacent to SNP sites, and especially, the accuracy of single-gene disease detection based on single or few cells can be improved.

The invention provides a kit, comprising a probe. The probe is fixed on a solid substrate or is free in a solution; the probe can specifically recognize a target area, wherein the target areas include one of the followings: at least one of the 163 genes shown in Table 1; or a CDS area of at least one gene in Table 1; or the upstream and downstream 10-20 bp area of a CDS area of at least one gene in Table 1. The invention also provides application of the kit, and a method and a system for detecting area target variation The kit and / or method and system can simply and conveniently obtain hereditary hearing impairment-related gene sequences at one time and high specificity, and accurately detect and analyze these related gene sequences, so that the detection and analysis results can assist the study of hereditary hearing impairment.

The invention relates to a specific primer and a test kit for detecting pathogenic variation of a deafness gene based on multiplex PCR and a high-throughput sequencing technology and application. Mosthereditary deafness can be subjected to high-efficiency, low-cost and comprehensive diagnosis; the specific primer and the test kit can also be used for newborn screening and early detection early treatment and can reduce matte caused by deafness, and the technical scheme for solving the problem is as follows: the multiplex PCR specific primer comprises 34 pairs of specific primers which are divided into two Pools, and Pool-1 comprises 18 pairs of primers, wherein the nucleic acid sequences are shown as SEQ ID NO: 1 to SEQ ID NO: 36; pool-2 comprises 16 pairs of primers, and the nucleotide sequence is shown as SEQ ID NO.37-68. According to the specific primer, the detection efficiency and accuracy can be effectively improved, the cost is reduced, the operation steps are simplified, and the specific primer is an innovation of specific primers for detecting the pathogenic variation of the deafness gene.