49 results about "BRAF Gene Mutation" patented technology

The invention discloses a nucleic acid probe, a liquid chip and a detection method that are used for the mutation detection of a BRAF gene exon 15. The liquid chip that is used for the mutation detection of the BRAF gene exon 15 comprises microspheres and a primer; wherein, the microspheres are coated with wild and mutant probes which are decorated with amino groups and aim at the BRAF gene exon 15; and the primer is used for augmenting a target sequence that is enriched in mutant sites needing to be detected by the BRAF gene exon 15 and has a biotin marker in the tail end. The detection method that is provided by the invention has quickness, accuracy, simple and convenient operation, and greatly improves the detection efficiency.

The invention discloses a kit for detecting colorectal cancer susceptibility gene mutation and a detection method thereof. The kit contains specific primers, universal primers, PCR (Polymerase Chain Reaction) mixing reagents, positive controls and magnetic beads aiming at PIK3CA, KRAS and BRAF gene mutation sites. The method comprises the following steps: extracting genome DNA of a sample to be detected, combining the specific primers with target template DNA sequences, amplifying a target area of the sample to be detected by the universal primers, purifying a library by the magnetic beads, performing high-throughput sequencing on the obtained library, and analyzing mutation conditions. The method and the kit disclosed by the invention can simultaneously detect a plurality of susceptibility genes, simplify experimental procedures and simultaneously improve the detection sensitivity, are suitable for performing early screening and postoperative diagnosis of the colorectal cancer, and provide guide for individualized medication.

The invention relates to a method and kit for detecting BRAF (Block Repeat Active Flag) mutation relevant to the curative effect of a molecular-targeting anti-cancer medicament, in particular relating to a fluorescence quantificational PCR (Polymerase Chain Reaction) detecting method and kit for detecting mutation in BRAF gene mutation hot spot regions and application thereof. According to the invention, mutation at special positions of BRAF genes is detected, the curative effect of the molecular-targeting anti-cancer medicament such as an EGFR (Epidermal Growth Factor Receptor) tyrosine kinase inhibitor, and the like can be forecasted, and furthermore, the individual medicament use schemes of patients with tumors are directed.

The invention discloses application of a BRAF gene detector to preparation of an ACTH type pituitary adenoma molecular pathological diagnosis and parting product. It is proved through tests that BRAF gene mutation is peculiar to ACTH type pituitary adenoma and not found in other types of pituitary adenoma, so that the BRAF gene mutation can serve as a specific molecular pathological diagnosis marker of the ACTH type pituitary adenoma. According to the result that a BRAF gene is mutant or not, the ACTH pituitary adenoma can be further divided into two subtypes, namely the BRAF mutant type and the wild type, the mean volume of the BRAF mutant type ACTH pituitary adenoma is small, the attack degree is low, and the midnight serum cortisol concentration is less than or equal to 22 ug / dl; the mean volume of the wild type ACTH pituitary adenoma is large, the attack degree is high, and the midnight serum cortisol concentration is greater than 22 ug / dl. Therefore, the different subtypes of the ACTH pituitary adenoma can be judged by detecting whether the BRAF gene is mutant or not, and different clinical treatment strategies are adopted in time accordingly.

The invention provides crRNAs for gene mutation of lung cancer-related molecular markers, a detection kit and a detection method thereof. The crRNAs comprise: a crRNA at the EGFR-T790M mutation site:as shown in any one of SEQ ID NO.16-17; a crRNA at the EGFR-L858R mutation site: as shown in any one of SEQ ID NO.18-19; a crRNA at the BRAF gene mutation site: as shown in any one of SEQ ID NO.20-21;a crRNA at the ALK gene mutation site: as shown in any one of SEQ ID NO.22-23; and a crRNA at the ROS1 gene mutation site: as shown in any one of SEQ ID NO.24-25. According to the invention, the 5 crRRNAs related to gene mutation targets of lung cancer-related molecular markers are designed, and the crRNAs are adopted in combination with a CRISPR-cpf1 system to carry out mutation detection. The method of the invention has the characteristics of simplicity, fast speed, high sensitivity and strong specificity.

The invention discloses a primer, a kit and a method for detecting BRAF (v-raf mourine sarcoma viral oncogene homolog B1) gene mutation. The primer for detecting the BRAF gene mutation comprises a forward amplification primer with a base sequence represented by SEQ ID NO:1, a backward amplification primer with a base sequence represented by SEQ ID NO:2 and a sequencing primer with a base sequence represented by SEQ ID NO:3, wherein the 5' tail end of the forward amplification primer and / or the backward amplification primer is provided with a biotin label. The method and the kit for detecting the BRAF gene mutation have the advantages of simplicity and rapidness for operation, high flux, low detection cost and the like, especially have good specificity, high sensitivity and high accuracy and have wide application prospect.

A method for fast detecting BRAF gene mutation with an allele RNA (ribonucleic acid) isothermal amplification method comprises the following steps: cell lysis treatment is performed; RNA reverse transcription treatment is performed; cDNA (complementary deoxyribonucleic acid) is generated and serves as an RNA amplification template; and RNA amplicon is quantified through regular monitoring for a fluorescence signal. According to the method, an isothermal reaction method is adopted for fixed-point specific detection of a V600E mutation site of the BRAF, the detection is performed at the isothermal temperature of 41 DEG C, test conditions are mild, chemical compounds for reaction are common, the operation is simple, the specificity is high, and only an instrument capable of detecting a fluorescent probe is required. A higher amplification coefficient is provided, a billion RNAs which are complementary with the target RNA are produced with two specific primers aiming at targeted RNA as well as three enzymes in 1.5 hours, and BRAF mutation information with as low as 0.1% of content is read through detection of the fluorescent probe and used for targeted drug use instruction for a tumor patient.

The invention belongs to the field of biotechnology, and specifically relates to a combination of primers for thyroid cancer detection, including DNA PCR primers and RNA PCR primers. The DNA PCR primers include ATM gene mutation detection primer pairs, BRAF gene mutation detection primer pairs, and HRAS gene mutation detection primers. Detection primer pair, NRAS gene mutation detection primer pair, PTEN gene mutation detection primer pair, RET gene mutation detection primer pair, TERT gene mutation detection primer pair, TG gene mutation detection primer pair, TP53 gene mutation detection primer pair, TSHR gene mutation detection Primer pair, TTN gene mutation detection primer pair; said RNA PCR primers include RET gene fusion detection primer pair, NTRK3 gene fusion detection primer pair, BRAF gene fusion detection primer pair. The invention can simultaneously detect DNA mutation and RNA fusion. The amount of sequencing data for a single sample detection is reduced to less than 0.2M reads, which greatly reduces the cost of sequencing. Since DNA mutation and RNA fusion are tested in the same system, the workload of operation is reduced.

The invention provides crRNAs for gene mutation of lung cancer-related molecular markers, a detection kit and a detection method thereof. The crRNAs comprise: a crRNA at the EGFR-T790M mutation site:as shown in any one of SEQ ID NO.16-17; a crRNA at the EGFR-L858R mutation site: as shown in any one of SEQ ID NO.18-19; a crRNA at the BRAF gene mutation site: as shown in any one of SEQ ID NO.20-21;a crRNA at the ALK gene mutation site: as shown in any one of SEQ ID NO.22-23; and a crRNA at the ROS1 gene mutation site: as shown in any one of SEQ ID NO.24-25. According to the invention, the 5 crRRNAs related to gene mutation targets of lung cancer-related molecular markers are designed, and the crRNAs are adopted in combination with a CRISPR-cpf1 system to carry out mutation detection. The method of the invention has the characteristics of simplicity, fast speed, high sensitivity and strong specificity.

The invention discloses application of a BRAF gene detector to preparation of an ACTH type pituitary adenoma molecular pathological diagnosis and parting product. It is proved through tests that BRAF gene mutation is peculiar to ACTH type pituitary adenoma and not found in other types of pituitary adenoma, so that the BRAF gene mutation can serve as a specific molecular pathological diagnosis marker of the ACTH type pituitary adenoma. According to the result that a BRAF gene is mutant or not, the ACTH pituitary adenoma can be further divided into two subtypes, namely the BRAF mutant type and the wild type, the mean volume of the BRAF mutant type ACTH pituitary adenoma is small, the attack degree is low, and the midnight serum cortisol concentration is less than or equal to 22 ug / dl; the mean volume of the wild type ACTH pituitary adenoma is large, the attack degree is high, and the midnight serum cortisol concentration is greater than 22 ug / dl. Therefore, the different subtypes of the ACTH pituitary adenoma can be judged by detecting whether the BRAF gene is mutant or not, and different clinical treatment strategies are adopted in time accordingly.

The invention discloses a primer set, a kit and a method for detecting BRAF gene mutation, and belongs to the technical field of gene detection. The primer set provided by the present invention comprises BRAF upstream primer and BRAF downstream primer; The nucleotide sequence of described BRAF upstream primer is as shown in sequence listing SEQ ID NO:1; The nucleotide sequence of described BRAF downstream primer is as sequence listing SEQ ID NO:2 shown. In addition, the kit provided by the present invention includes PCR amplification reaction reagents, positive quality control products and negative quality control products, primer mixture, primer-probe mixture and standard curve equation. The method for detecting BRAF gene mutation provided by the present invention has the characteristics of high efficiency, simplicity, intuition, high sensitivity, etc., and can quantitatively analyze the mutation frequency of BRAF gene.

The invention discloses a reagent storage device for detecting BRAF gene mutation. In the invention, the cooling liquid is used as the temperature maintaining medium of the reagent tube, which can effectively improve the temperature maintaining stability of the reagent tube. The cooling liquid in the liquid exchange and cooling components is monitored in real time. When the liquid is exchanged synchronously, the cooling liquid in the circulating pipeline can be supplemented into the liquid exchange tank by the suction of the micro suction pump, and the Part of the cooling liquid in the liquid exchange tank is discharged, so that the cooling capacity of the cooling liquid in the liquid exchange tank can be smoothly improved to reach the initial level. Push the push plate to move, the push plate can push out the liquid in the liquid exchange tank, and the distance between the top of the push plate and the top of the liquid exchange tank can ensure that the push plate is automatically reset after the liquid exchange is completed, which improves the liquid exchange rate. Efficiency ensures the effectiveness of the reagents, thereby ensuring the accuracy of the detection.