37 results about "Gene mutation analysis" patented technology

The invention discloses primers for detecting ApoE gene polymorphism, a kit and a PCR (polymerase chain reaction) method for the primers or the kit. The primers include two sets of primers for detecting an rs429358 site and an rs7412 site respectively; the first set of primers include a mutant-type specific upstream primer of the rs429358 site, a wild-type specific upstream primer of the rs429358 site and a first downstream primer shared by the mutant-type specific upstream primer and the wild-type specific upstream primer of the rs429358 site; the second set of primers include a mutant-type specific upstream primer of the rs7412 site, a wild-type specific upstream primer of the rs7412 site and a second downstream primer shared by the mutant-type specific upstream primer and the wild-type specific upstream primer of the rs7412 site. The kit has the advantages of simplicity, quickness, accuracy and low price for detection and the like, and provides a powerful tool for scientific research and clinical analysis of ApoE gene typing and gene mutation.

The invention relates to a high-sensitivity kit for quantitatively detecting EGFR (Epidermal Growth Factor Receptor) gene mutation and application thereof. The application comprises the following steps: respectively designing a forward primer and a backward primer in a sequence with the length of 50-100bp in a specific amplification gene mutation site area aiming at a to-be-detected gene template, designing the mutation sites at corresponding positions of the forward primer, and designing competitive Block oligonucleotides for inhibiting non-mutant amplification nearby the mutation sites. The detection is performed by utilizing the specificity aiming at a probe of the amplification product, and the gene mutation conditions are obtained by detecting the detectable markers on the probe. The invention provides a gene mutation analysis method which is stable in result, high in sensitivity and high in repeatability.

The invention provides a kit for detecting age-related macular degeneration disease, which can be used for early monitoring and detection of AMD high-risk groups, implement early prevention and treatment, and reduce their visual impairment. The kit of the present invention targets the 512G→A mutation of the HTRA1 gene and its related sites/genes, has a high correlation with AMD, and cooperates with the analysis of the CHF gene mutation to improve the accuracy of detection.

The invention relates to a gene detection kit for prognosing gastric cancer metastasis. The kit comprises a DNA library building kit, wherein the DNA library building kit comprises a high-risk gene probe and a low-risk gene probe; the high-risk gene probe comprises CDH1, CDH2, SNAIL, SLUG, MUC4, MUC6, PRSS3, USP6, MLH1, MSH2, MSH6, PMS2, TGFBR2, MMP2, MMP9, BRCA1, BRCA2, PALB2, ATM, ATR, MUTYH, EMSY, ERCC4, RAD51, PARP1 and XRCC1; the low-risk gene probe comprises ATRX, BRIP1, FANCA, FANCB, FANCC, FANCD2, FANCE, FANCF, FANCG, FANCI, FANCL, FANCM, FANCO, FANCP, MDM2, MDM4, MLH1, NPM1, PP2R1A, PRKDC, RAD50, STAG2, XRCC5 and CRCC6. The invention further discloses a use method of the kit. The use method comprises the following steps: extracting cfDNA in a blood sample; building a library for the cfDNA through the DNA library building kit, and then sequencing the DNA to obtain a gene overall length sequence; carrying out gene mutation analysis on the gene overall length sequence.

The invention discloses an algorithm for the automatic generation module of the HGVS (Human Geome Variation Society) name of gene mutation. The algorithm comprises the following elements: 1) describing a mutation site by a ''gene(R/M)Sz/c'' format, wherein the meaning of the gene(R/M)Sz/c is a gene name, one segment of sequence after a (mutation site reference sequence/ mutation site mutation sequence) mutation site and whether the mutation site is a homozygotic type or a hybrid type, and the three parts are separated by a space so as to describe mutation discovered by sequencing; 2) the format is input into a mutation analysis system, a gene information database, a gene inheritance pattern way database, an HGMD database and the like are called through an HGVS calculation program for calculation; and 3) the HGVS name of the gene mutation and relevant information based on the HGVS information are obtained. Through tests, a human gene mutation system based on the algorithm can analyze of mutation of more than 6000 human genes so as to meet the analysis requirements of gene mutation including common genetic diseases, cancers and the like.

The invention relates to a circulating tumor DNA detection system for screening existence of postoperative minimal residual focuses of colorectal cancer patients and predicting recurrence risk. The detection system comprises a colorectal cancer tissue gene mutation screening module, a plasma free DNA gene mutation analysis module and a circulating tumor DNA state judgment module. The system utilizes the same next-generation sequencing gene combination panel to detect primary tumor tissue and plasma free DNA, and considers all mutations detected in the primary tumor tissue of a patient instead of being limited to individual gene mutations, so that the system is more comprehensive. The system can be applied to dynamic and real-time monitoring of circulating tumor DNA, and can be used for evaluating the residual condition of minimal residual lesions after colorectal cancer radical operation, predicting the recurrence risk and guiding the postoperative treatment decision.

The invention relates to the field of gene engineering and cytology, in particular to an AAV receptor knocking out method, an HEK293 cell strain with an AAV receptor being knocked out and an application. The knocked-out AAV receptor is KIAA0319L. The knocking out method comprises the following steps: establishing a sgRNA carrier of a targeted AAVR genome sequence; carrying out transfection for a 293 T-cell, and culturing to obtain each clonal cell; carrying out Suveyor gene mutation analysis test; selecting AAVR mutant monoclone; then selecting an AAVR mutant cell strain with high yield of AAV; identifying a mutation condition of other potential mutant site; and enlarging the cell strain without other mutant sites, and establishing a library for production. The final yield of the AAV produced by the HEK293 cell strain of the knocked-out AAV receptor is about 50 percent higher than the yield of the conventional HEK293.

The invention discloses a primer for detecting BRAF gene V600E mutation sites, and a PCR method of the kit. The primer comprises a wild type specific forward primer, a mutant type specific forward primer, and a reverse primer shared by the wild type specific forward primer and the mutant type specific forward primer, wherein the wild type specific forward primer has a sequence shown as SEQ No.17; the mutant type specific forward primer has a sequence shown as SEQ No.14; and the shared reverse primer has a sequence shown as SEQ No.16. The kit has the advantages of simple detection, rapidness, accuracy, low price and the like and provides a powerful tool for scientific research and clinical detection of BRAF gene V600E mutation sites and gene mutation analysis.

The invention discloses a tumor neoantigen-based T cell sorting preparation technology and application thereof in the technical field of molecular biology, and the T cell sorting preparation technology based on tumor neoantigen and application thereof comprise the following steps: S1: collecting a tumor specimen, analyzing tumor gene mutation through gene sequencing, and collecting tumor tissues of a patient by means of surgery, puncture, intervention and the like; performing sequencing and detecting gene mutation through methods such as full-exon sequencing and probe recovery method sequencing, S2: analyzing the tumor neoantigen according to the gene mutation, and analyzing the tumor neoantigen according to information such as the gene mutation, HLA affinity and gene expression. The T cell sorting preparation technology based on the tumor neoantigen and the application of the T cell sorting preparation technology are characterized in that NRT cells are sorted out based on tumor neoantigens, pure NRT cells are obtained, and preparations are made for subsequent cell treatment in the aspects of safety, cost and effectiveness.

The invention discloses a kit for detecting drug sensitivity of rectal cancer cells based on gene mutation analysis, which comprises the following substances: (1) a rectal cancer cell drug reagent, (2) a magnetic nanoparticle indicating liquid A with a green fluorescent protein labeling a wild P53 protein antibody for surface modification, (3) a magnetic nanoparticle indicating liquid B with a redfluorescent protein labeling an anti-mutation P53 protein antibody for surface modification and (4) a substrate medium and a dilution buffer. The sensitivity of a patient to the drug is measured through simultaneous calculation of the expression levels of the wild P53 gene protein and the mutant P53 gene protein, the individualization is more prominent, the detection result is more accurate, a sensitive drug is screened for the treatment of the disease, and an optimal chemotherapy regimen is selected.

The invention discloses a gene mutation analysis judgment method based on a real-time fluorescence quantification PCR (polymerase chain reaction) platform. A fluorescent quantitation PCR amplification result of a target gene is used for analysis judgment. The analysis judgment method comprises steps as follows: step one, after a PCR experiment ends, an experimental result is subjected to CT (computerized tomography) value calculation, and a CT value is judged to determine whether the experiment condition is continuously analyzed; step two, internal control is operated, and whether a DNA (deoxyribonucleic acid) sample is qualified is judged according to the internal control result; step three, a positive quality control product is operated, and whether an experiment system is qualified is judged according to the CT value of the positive quality control product; and step four, if the CT value is qualified through check analysis, a Tm calling program is operated, and whether the sample has mutation is judged through melting curve analysis and CT value analysis. According to gene mutation analysis judgment method, a PCR result is clearly and objectively judged, sample gene types are judged according to amplification data and a melting curve, and the misjudgment rate is low.

The invention relates to a true and false gene mutation analysis method based on high-throughput sequencing and application, and belongs to the technical field of bioinformatics. The true and false gene mutation analysis method comprises the following steps: acquiring difference sites in reference sequences of homologous true genes and false genes; comparing the NGS sequencing data with the difference sites to respectively obtain the true gene reads number and the false gene reads number corresponding to the same difference site, taking the ratio of the true gene reads number to the false genereads number of the same difference site as a judgment index, and carrying out mutation analysis and judgment on the true gene according to a predetermined strategy. According to the kit, mutation oftrue and false genes can be preliminarily screened, genes which possibly have problems are found out, and judgment is carried out in combination with clinical actual conditions. MLPA or sanger sequencing experiments carried out on genes one by one are avoided, and the experiment cost and time are greatly saved.

qRT-PCR primers capable of detecting EML4-ALK gene variant based on circulating tumor cells at a more sensitive detection limit than a conventional method. Also disclosed is a lung cancer patient screening method which is capable of detecting EML4-ALK gene variant using circulating tumor cells even in a lung cancer patient on whom ALK-FISH testing has been difficult to perform, due to difficulty in collecting a solid lung cancer tissue sample, and is able to determine whether an anticancer drug targeting the EML4-ALK gene variant may be applied to the lung cancer patient.