24 results about "Hotspot mutation" patented technology

The invention discloses an anti-TP53 mutation T cell receptor and application thereof, the T cell receptor comprises specific alpha chain and beta chain variable domains, and the complementary determining region (CDR) sequence is shown as SEQ ID NO: 9-14. The TCR has the core advantage that the TCR has excellent broad-spectrum recognition capability, can target six different amino acid substitutions (A, G, I, N, S and T) at the R249 site, and effectively deals with tumor heterogeneity and mutation difference between patients. Aiming at high-frequency HLA-B * 07: 02 alleles in people, the TCR lays a foundation for developing TCR-T cell therapy covering a wide range of people, and has great clinical application value and market potential in treatment of various solid tumors carrying TP53 R249 hotspot mutation, such as liver cancer.

The invention relates to a detection kit for combined methylmalonemia, which is a cb1C type kit and comprises a specific primer pair and a probe combination for detecting at least one mutation site of MMACHC, the mutation site is selected from c.609Ggt; a is c.567 dupT, c.658660delAAG, c.482Ggt, and c.482Ggt; a, c.1Agt; g, c, 80Agt; g, c, 217Cgt; 315Cgt, T, c.315Cgt; g and c.394 Cgt; t). The MMACHC gene hotspot mutation region detection kit based on the real-time fluorescent quantitative PCR technology can detect multiple mutation sites on the MMACHC gene at the same time, detection of three genotypes of wild type, homozygous mutant type and heterozygous mutant type on each mutation site is completed, the gene detection requirements of clinical combined type MMA are effectively met, the detection time is shortened, and the detection efficiency is improved. The mutation site coverage rate of CblC defective methylmalonic acidmia combined with homocysteinemia on the MMACHC gene is increased to 93.28%, mutation hot spots of most people are covered, and various types of mutation conditions in hot spot mutation areas can be rapidly, accurately and sensitively detected.

The invention relates to a kit for absolutely and quantitatively detecting KCNJ5 hotspot mutation based on a ddPCR technical platform and a detection method of the kit. The kit comprises an upstream primer KCNJ5-451F, a downstream primer KCNJ5-503R, mutant probes KCNJ5-451P-MUT, mutant probes KCNJ5-451P-MUT, mutant probes KCNJ5-503P-MUT, wild probes KCNJ5-451P-WT, wild probes KCNJ5-451P-WT and wild probes KCNJ5-503P-WT. The invention further relates to a kit for absolutely and quantitatively detecting KCNJ5 hotspot mutation based on the ddPCR technical platform and a method for absolutely and quantitatively detecting KCNJ5 hotspot mutation based on the ddPCR technical platform. The kit is simple, convenient and rapid, absolute quantitative detection of KCNJ5 gene hotspot mutation can be completed only through a single-tube reaction system, the kit has good sensitivity and repeatability, the detection result is accurate and reliable, an important gene marker result is provided for prediction of the postoperative curative effect of an APA patient, and the kit has important clinical application value.

The invention discloses a primer group and a kit for detecting spinal muscular atrophy related genes and a kit using method, the primer group comprises an amplification primer group and an extension primer group, primer nucleotide sequences in the amplification primer group are as shown in SEQ ID NO: 1-14, and primer nucleotide sequences in the extension primer group are as shown in SEQ ID NO: 15-26. The kit provided by the invention comprises the primer group, a PCR reaction reagent, an SAP reaction reagent and an extension reaction reagent, and can simultaneously detect SMN1 and SMN2 gene copy numbers, 7 hotspot mutations in people and c.* 3 + 80Tgt closely related to silent carriers in one reaction; the qualitative detection of the G site has a wider detection range for spinal muscular atrophy. Meanwhile, the amount of sample templates required by nucleic acid detection of the kit is small, the detection limit of the nucleic acid templates can be as low as 1 ng / mu L, and the kit has good sensitivity, specificity, stability and accuracy and can meet the current needs of screening patients with spinal muscular atrophy and silent carriers and prenatal diagnosis of infants.

The present invention provides a primer combination, a kit, a detection system, a detection method and an application for detecting pathogenic gene mutations of familial Alzheimer's disease and APOE genotypes by means of constant temperature multiplex rapid detection. The method of the present invention uses the amplification refractory mutation system technology combined with the nicking endonuclease nucleic acid detection reaction isothermal amplification system to perform multiplex amplification on 14 sites including 12 hot spot mutations and 2 risk sites of the Alzheimer's disease pathogenic gene, and at the same time combines capillary electrophoresis for product analysis to perform genotyping detection.

The application discloses a method for detecting glioma chromosome abnormalities based on targeted sequencing, and belongs to the technical field of biological medicine. The method first acquires the allele frequency of a to-be-detected sample at preset SNP sites (covering 1p, 1q, 19p, 19q, chromosome 7 and chromosome 10), and then calculates and determines whether specific chromosome arms or chromosomes have loss of heterozygosity. Meanwhile, the copy number of the region where each SNP site is located is calculated based on the sequencing depth, and the total copy number of the above-mentioned chromosomes is obtained by integration. Finally, the loss of heterozygosity determination result and the chromosome copy number information are comprehensively combined, so that the simultaneous identification of 1p / 19q co-deletion, gain of chromosome 7 (+7) and deletion of chromosome 10 (-10) is realized. The method does not require paired samples, can accurately quantify the copy number, avoid false positives, and only needs to detect part of the SNP sites, that is, can be combined with hot spot mutation detection, thereby saving cost and improving detection efficiency.

Provided and featured are novel recombinant T cell receptors (TCRs) that target a mutated tumor suppressor TP53. Also provided are cells comprising such TCRs, and methods of using the recombinant TCRs and cells expressing the recombinant TCRs for treating cancers associated with mutated TP53.

The invention provides a primer for detecting Exon10 hot spot mutation of an MPL gene and application of the primer, the primer for detecting the Exon10 hot spot mutation of the MPL gene comprises amplification primers and sequencing primers, the sequences of the amplification primers are 5 '-AGTAGGGGCTGGCTGGATGA-3' and 5 '-ACCTGACGCAGATTGGATGG-3', and the sequences of the sequencing primers are 5 '-AGTAGGGGCTGGGATGG-3', 5 '- And the sequences of the sequencing primers are 5 '-GGCCGAAGTCTGACCCTT-3' and 5 '-GCGGTATAGTGGGCGTGTT-3'. The PCR amplification primers and the sequencing primers are separated, one pair of amplification primers is firstly used for PCR amplification, and the other pair of sequencing primers is used for sequencing, so that a single peak with a better sequencing effect is obtained, the mutation condition is accurately judged, and the sensitivity is high.

The application belongs to the technical field of gene detection and molecular diagnosis, and particularly relates to a kit for detecting a pathogenic gene of phenylketonuria and application thereof. The kit comprises a primer pair for amplifying specific exons and splicing regions of a phenylalanine hydroxylase gene, and specific probes for detecting hot spot mutations and deletion / repetition variations of the gene. The application can detect genetic variations related to phenylketonuria in one time, quickly and accurately by combining optimized multiplex polymerase chain reaction with high-throughput sequencing or gene chip technology, and covers various known hot spot mutations and copy number variations including c.1222C>T, c.1068-11G>A, c.728G>A and c.1162G>A. The kit has high detection sensitivity and strong specificity, and is suitable for positive recall diagnosis of neonatal phenylketonuria screening, genetic diagnosis of suspected patients, carrier screening and prenatal diagnosis, and provides an efficient tool for precise prevention and control of phenylketonuria.

Provided and featured are novel recombinant T cell receptors (TCRs) that target a mutated tumor suppressor TP53. Also provided are cells comprising such TCRs, and methods of using the recombinant TCRs and cells expressing the recombinant TCRs for treating cancers associated with mutated TP53.

The present invention discloses a primer-probe combination for detecting KRAS, NRAS, BRAF and PIK3CA gene mutations and its application. The primer-probe combination includes primers and probes for detecting KRAS, NRAS, BRAF and PIK3CA gene mutations. The present invention designs a primer-probe combination for detecting KRAS, NRAS, BRAF and PIK3CA gene mutations, and uses digital PCR to qualitatively detect 14 hotspot mutations of KRAS, NRAS, BRAF and PIK3CA genes in tumors, with good specificity and sensitivity, simple operation and low cost.

The present invention provides and features novel recombinant T cell receptors (TCRs) that target mutated tumor repression factor TP53. Also provided are cells comprising such TCRs, as well as methods of using the recombinant TCRs and cells expressing the recombinant TCRs to treat cancers associated with mutated TP53.

The invention relates to the technical field of tumor gene mutation, in particular to a tumor gene mutation interpretation method based on an NGS platform, and the method comprises the steps: obtaining a mutation detection result of a to-be-analyzed sample, the mutation detection result comprising at least one candidate mutation and mutation data corresponding to the candidate mutation; judging whether the candidate mutation exists in a preset list or not according to the mutation detection result; if the candidate mutation exists in the preset list, generating an interpretation result corresponding to the candidate mutation according to the list type of the preset list; if the candidate mutation does not exist in the preset list, determining a hotspot mutation type and a sample quality type of the candidate mutation of the to-be-analyzed sample according to a mutation detection result; and based on the hotspot mutation type, the sample quality type and the mutation data, generating an interpretation result corresponding to the candidate mutation. The method has the advantages that the standardization level and clinical applicability of tumor gene mutation interpretation can be improved, and meanwhile the false positive misjudgment rate and omission ratio are effectively reduced.

The invention discloses a primer combination, a kit and a library construction method for detecting POLE gene multi-hotspot mutation, the primer combination is used for targeted amplification of POLE gene mutation sites, and the primer combination comprises P286R, S297F, M295R, F367S, D368Y, V411L, L424I, P436R, M444K, S459F and A456P. The kit can detect 11 mutation sites of the POLE gene at the same time, directly reflects mutation conditions of the sites, and is used for endometrial cancer POLE molecular typing.

The invention belongs to the field of biological medicines, and provides a device for predicting disease recurrence based on an ultrahigh-sensitivity mutation load, an MRD gene detection panel covering 28 hotspot mutation regions of acute myelogenous leukemia is designed by integrating famous tumor public databases such as COSMIC, a quantitative blocker displacement amplification QBDA technology is combined, and the disease recurrence is predicted. According to the method, ultra-low abundance mutation with the variation allele frequency lower than 0.01% can be detected, a set of device for predicting relapse in advance based on ultra-high sensitivity mutation load change in the complete remission period of an acute myelogenous leukemia patient is developed, and the important clinical application value is achieved.

The present invention relates to an in vitro method for identifying and / or characterizing one or more mutations in a hotspot mutation sequence of at least one ESR1 target fragment from a DNA sample, with a drop-off digital polymerase chain reaction (PCR).

The present invention discloses an sgRNA guide sequence targeting RP1L1 gene and its application, which relates to the field of genetic engineering technology. The technical key points are as follows: the nucleotide sequence of the sgRNA is as shown in SEQ ID NO: 1, the sgRNA is added with a backbone sequence and sgRNA1 is prepared by 2'-O-methylation modification and thiolation modification, the nucleotide sequence of the sgRNA1 is as shown in SEQ ID NO: 2, and the backbone nucleotide sequence is as shown in SEQ ID NO: 3. The sgRNA targeting the hotspot mutation c.133C>T of the RP1L1 gene provided by the present invention can not only accurately target and edit the mutation sequence, but also mediate homologous recombination to correct the mutation sequence to a normal sequence. This sgRNA, as an efficient guide sequence, can be applied in multiple fields such as the study of pathogenic mechanisms related to the RP1L1 gene, the construction of cell and animal models, and drug development.

Described herein are methods and compositions for analyzing nucleic acid sequences. Amplicon sequencing is particularly useful for genome targeting and detection of hot-spot mutations, copy number variations, gene fusions, InDels and single-nucleotide polymorphisms (SNPs). However, for applications such as personalized cancer monitoring, wide tracking of a plurality of variants requires use of a primer sequences of extraneous length. Methods and compositions are described herein that support reduction of the length of primers with benefits including creased purity, error reduction, reduced manufacture time and savings in cost and time if a shared primer could be applied.

The invention discloses a kit for detecting hot spot mutation Q61R / Q61K of a human NRAS gene based on an LNA-AS qPCR principle and application of the kit. The kit comprises two forward modified primers, one reverse universal primer and one Taqman detection probe which are used for detecting hot spot mutation. The kit disclosed by the invention adopts an LNA-AS Taqman qPCR method, and has the remarkable advantages of low unit cost, short time consumption and simplicity in operation on the basis of ensuring accuracy. The kit disclosed by the invention has a very good application prospect for detecting two point mutations of the human NRAS gene in early screening and accompanying diagnosis of lung cancer and colorectal cancer.

Techniques for determining prognostic classifications of pathologies are described. An example method includes determining features of a sample from a subject, generating input data indicating the features, and generating, based on the input data and a predictive model, a prognostic classification of the subject. The features include one or more of: a MMRD probability score of the sample, a copy number state of the sample, a presence of one or more pathogenic variants, a fraction unstable score, a mutation signature, a TMB score, the presence of a hotspot mutation, or the presence of an aneuploid event. The MMRD probability score is indicative of a pathogenic variant in at least one MMR gene, a methylation status of the at least one MMR gene, or a methylation status of one or more promotors associated with the at least one MMR gene.

The invention discloses a multi-gene mutation tumor new antigen polypeptide algorithm, the specific sites of the mutated antigen targets are genes APRT p.E141V, KCTD10 p.Y172N, PABPC1 p.L218V, ADAM28 p.C315G, ARF3 p.G6V, KLHL2 p.E88G, CNOT11 p.S148R, C5orf42 p.Q1002L, ABLIM1 p.C190Y, BCLAF1 p.N629S, PABPC1 p.K312RfsTer10 and KRAS p.G12D, the 12 gene mutations cover hotspot mutation, low-frequency mutation and rare mutation, and the antigen polypeptide can be synthesized on a large scale and is used for subsequent PABPC1 p.L218V and a more effective personalized treatment scheme can be established for the patient.

This application relates to the field of tumor gene mutation technology, and in particular to a tumor gene mutation interpretation method based on an NGS platform. The method includes acquiring mutation detection results of a sample to be analyzed, the mutation detection results including at least one candidate mutation and its corresponding mutation data; determining whether the candidate mutation exists in a preset list based on the mutation detection results; if the candidate mutation exists in the preset list, generating an interpretation result corresponding to the candidate mutation based on the list type; if the candidate mutation does not exist in the preset list, determining the hotspot mutation type and sample quality type of the candidate mutation in the sample to be analyzed based on the mutation detection results; and generating an interpretation result corresponding to the candidate mutation based on the hotspot mutation type, sample quality type, and mutation data. This application can improve the standardization level and clinical applicability of tumor gene mutation interpretation, while effectively reducing the false positive rate and the missed detection rate.

The invention discloses a primer combination, a kit and a method for BCR-ABL1 fusion gene quantification and ABL1 kinase region hotspot mutation synchronous detection, and belongs to the technical field of molecular diagnosis and gene detection. On the basis of a detection strategy of RT-PCR + capillary electrophoresis integration, by optimizing primer design and a detection process, the problems of low detection flux, complicated operation, high cost and the like in the prior art are solved, 13 fusion subtypes of BCR-ABL1 can be identified at the same time, more than 90% of clinical related drug-resistant mutation sites in an ABL1 kinase region can be covered, and the application prospect is wide. The kit has the technical advantages of integrated detection process, high specificity, high detection sensitivity, rapidness, high efficiency and the like, and is particularly suitable for clinical popularization and application.

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.