67 results about "Brca1 gene" patented technology

The invention relates to the technical field of gene detection, and provides a method and a primer for detecting mutation sites of all exon sequences of human BRCA1 and BRCA2 genes. The specific primers include forward and backward primers for amplifying all 24 exons covering the BRCA1 gene, and base sequences of the primers are as shown in SEQ ID NO: 001-062, as well as forward and backward primers for amplifying all 27 exons covering the BRCA2 gene, and base sequences of the primers are as shown in SEQ ID NO: 063-142. By virtue of a Sanger sequencing method, the method and the primer disclosed by the invention can be used for detecting the mutation of all exons of the BRCA1 gene and the BRCA2 gene and can be used for extending all exons of the entire BRCA1 and BRCA2 genes, covering all to-be-detected mutation sites. The method and the primer disclosed by the invention are quite high in specificity and accuracy, simple to operate and low in cost, and can be used for greatly enhancing the risk assessment of hereditary breast cancer and effectively reducing the onset risk of the breast cancer.

The invention discloses a mutant gene group for mammary cancer risk assessment and a detection kit thereof. The mutant gene group is a mutant gene set comprising the following seven genes: BRCA1 gene, BRCA2 gene, TP53 gene, MLH1 gene, MLH3 gene, MSH3 gene and CDH1 gene. The mutant gene group of the seven genes totally carries 25 mutant sites disclosed as Table 3. The invention also discloses a detection kit of the mutant gene group for mammary cancer risk assessment, which comprises probes or primers for detecting the mutant genes in the mutant gene group. The detection kit disclosed by the invention has the advantages of high detection efficiency, high detection speed, high detectable rate and the like.

The invention discloses a primer, a method and a kit for detecting mutation sites of human BRCA1 and BRCA2 gene all-coding sequences. The primer comprises forward and reverse primers which can amplifyfor covering all the 24 coding zones of BRCA1 gene and of which base sequences are shown as SEQ ID NO:3-52. The primer further comprises forward and reverse primers which can amplify for covering allthe 27 exon coding zones of BRCA2 gene and of which base sequences are shown as SEQ ID NO: 53-134. A detection method based on NGS technique comprises the following steps: combining a specific primerwith a target template DNA sequence; adopting a universal primer for amplifying a to-be-detected sample target area; purifying a library by using a magnetic bead; performing high-throughput sequencing on the acquired library and analyzing mutation of BRCA1 and BRCA2 genes. The primer, method and kit disclosed by the invention can cover all the to-be-detected mutation sites; uniformity is high; experimental procedures can be simplified; detection sensitivity is promoted; a guidance is supplied for selective medication of patients suffering from familial breast cancer and ovarian cancer as wellas for genetic predisposition; morbidity risk is effectively reduced.

Large deletions have been identified in the BRCA1 gene in patients. The large deletions predispose the patients to breast cancer and ovarian cancer. Thus, methods for detecting the genetic variants are provided which can be used in detecting a predisposition to cancer.

The invention relates to a fluorescence quantitative PCR (Polymerase Chain Reaction) diagnostic kit for rapid detection of mRNA expression level of a BRCA1 gene. The kit comprises BRCA1gene primer, a reference gene GAPDH primer and a Taqman fluorescence probe. The BRCA1 is an important cancer suppressor gene, and proteins coded by the BRCA1 gene play an important role in DNA damage and repair. A fluorescence quantitative PCR with high sensitivity and specificity is employed to detect the mRNA expression level of BRCA1; and a detection result has substantially increased specificity and sensitivity. The kit provides a novel rapid simple gene diagnostic technique for whether a clinical malignant tumor patient should use platinum chemotherapeutics and antimicrotubular medicaments.

The invention discloses a multiple PCR kit for detecting the eleventh exon mutation of breast cancer susceptibility gene and a preparation method thereof. The PCR amplification primers of the kit are composed of primers pair 1 which consist of SEQIDNO:1 and SEQIDNO:2 and primers pair 2 which consist of SEQIDNO:3 and SEQIDNO:4. The multiple PCR kit of the invention combines the SSCP technology with the DNA silver staining and can simultaneously screen the mutation conditions of the eleventh exon of BRCA1 gene and the eleventh exon of BRCA2 gene. The invention has the advantages of strong sensitivity, high accuracy, simple and convenient property, etc. and has important significance on the early detection and the forecast of the breast cancer.

The invention discloses a PCR (Polymerase Chain Reaction) kit for detecting BRCA (Breast Cancer) genic mutation, which is characterized by comprising a plurality of primer pairs used for specifically amplifying a BRCA gene targeting sequence, wherein each primer pair contains a continuous nucleotide sequence formed by at least 15 continuous nucleotides in the 2nd exon of a BRCA1 gene or the 20th exon of the BRCA1 gene or the 11th exon of a BRCA2 gene; the 2nd exon of the BRCA1 gene has a continuous nucleotide sequence of SEQ ID No:1; the 20th exon of the BRCA2 gene has a continuous nucleotide sequence of SEQ ID No:2; and the 11th exon of the BRCA2 gene has a continuous nucleotide sequence of SEQ ID No:3. The kit completes the judgment on the sample genotype by adopting a saturation probe and a high resolution dissolution curve analyzing technology, thereby providing the guidance for medication selection and genetic predisposition of familial breast cancer and ovarian cancer.

The present invention discloses a construction method of a tumor BRCA1 / 2 gene variation library for high-throughput sequencing detection, wherein the tumor BRCA1 / 2 gene variation library covers the mutations at all the exon regions of the human BRCA1 / 2 gene. According to the present invention, the complete exon sequences of the BRCA1 gene and the BRCA2 gene are subjected to single-tube amplification to rapidly complete the construction of the library, wherein the construction time of the whole library is only 2-3 h, and the manual time is only 15-30 min; and by combining the obtained library and the high-throughput sequencing platform, the difficult problem that the detection of the complete exon sequence is required on the basis of the small amount of the clinical samples in tumor diseases in the clinic can be effectively solved, and the cost is low.

Large deletions have been identified in the BRCA1 gene in patients. The large deletions predispose the patients to breast cancer and ovarian cancer. Thus, methods for detecting the genetic variants are provided which can be used in detecting a predisposition to cancer.

The invention discloses a BRCA1 / 2 gene variation deciphering database and a constructing method thereof. The database comprises Chinese population BRCA1 / 2 gene variation histological information, Chinese population BRCA1 / 2 gene variation frequency and Chinese population BRCA1 / 2 gene variation deciphering result. According to the BRCA1 / 2 gene variation deciphering database according to the invention, the Chinese population BRCA1 gene and BRCA2 gene variation site information and related information of variation frequency in Chinese population are firstly collected and processed comprehensively.Furthermore a deciphering result which aims at the Chinese population BRCA1 / 2 gene variation is included. The database is established for aiming at the Chinese population characteristic. Therefore the variation characteristic of the Chinese population BRCA1 / 2 gene can be presented in a relative comprehensive manner. Therefore the deciphering result is more suitable for the Chinese population.

Methods for identifying functional allele profiles of a given gene are disclosed. Functional allele profiles comprise the commonly occurring alleles in a population, and the relative frequencies at which such alleles of a given gene occur. Functional allele profiles are useful in treatment and diagnosis of diseases, for genetic and pharmacogenetic applications and for evaluating the degree to which the gene(s) are under selective pressure.

The invention relates to a gene probe composition and a kit for detecting epithelial ovarian cancer. The gene probe composition comprises a c-myc gene probe, an Rb1 gene probe, a Chk2 gene probe, a p53 gene probe and a BRCA1 gene probe. Another objective of the invention is to provide a kit for fluorescence in situ hybridization (FISH) detection of the epithelial ovarian cancer, and the kit comprises the gene probe composition as described above. The kit can detect 5 kinds of genes in a same sample even a single tumor cell of the epithelial ovarian cancer at the same time, and thereby substantially improving detection capability and efficiency. The invention further provides an optimal sample selection strategy and a slide preparation method for an ovarian cancer FISH research.

The invention provides a fluorescent biosensor to detect BRCA1 gene and a preparation method thereof. A composite for detecting BRCA1 gene comprises biotin-modified DNA Walker chains (DWTs), a protective probe (PP), biotin-modified substrate chain DNA (SDs), streptavidin-modified magnetic beads (SMBs), a FAM-modified fluorescent signal chain P, a Dabcyl-modified L chain, an S chain and an auxiliary chain A. The fluorescent biosensor and detection system to detect BRCA1 are constructed successfully herein; BRCA1 measurement with the fluorescent biosensor applied herein shows high sensitivity, good stability and good reproducibility. Compared with the prior art, the fluorescent biosensor has high detection speed, good sensitivity, good operational simplicity and convenience, short detectioncycle, good specificity, low false positive rate and low false negative rate. The fluorescent biosensor is applicable to the measurement of actual samples and the like and is expectedly applicable asa sensor having actual application value.

The present invention relates generally to the field of human genetics, and more specifically to the detection of a specific type of germline mutations in the BRCA1 gene, which will predispose to breast and ovarian cancer. In addition, the invention relates to the molecular genetic mechanism that may have mediated the genesis of these mutations, in particular the role of Alu repetitive DNA elements present in the intronic regions of BRCA1. The invention further relates to somatic mutations of this type in the BRCA1 gene in human breast and ovarian cancer, and their use in the diagnosis and prognosis of human breast and ovarian cancer. The invention more particularly relates to the screening of this type of BRCA1 mutations in human genomic DNA, which are useful for the diagnosis of inherited predisposition to breast and ovarian cancer.

Systems and methods for determining whether a cancer patient may respond to PARP inhibitor and / or platinum chemotherapy based on identifying exon excision variants in the BRCA 1 gene are provided. Exon excision variants may encode a hypomorphic BRCA1 protein. The cancer patient may be a breast cancer patient or an ovarian cancer patient. The patient may have any cancer in which exon deficiency in the BRCA1 gene contributes to resistance to PARP inhibitor or platinum therapy.

The invention provides methods for identifying mutations, such as single nucleotide polymorphisms (SNPs), within breast and ovarian cancer associated genes that modify the binding efficacy of microRNAs (miRNAs). In a preferred embodiment, methods of the invention identify a SNP that decreases expression of the BRCA1 gene by increasing or decreasing the binding efficacy of at least one miRNA. Alteration of miRNA binding to BRCA1 by the introduction of SNPs within miRNA binding sites modulates or decreases BRCA1 expression, ultimately leading to the unregulated cell proliferation of a breast or ovarian cancer cells.

The invention provides a BRCA1 / 2 genovariation combined detection kit and belongs to the field of genovariation detection. The kit comprises a BRCA1 gene primer group, a BRCA2 gene primer group, polymerase, a PCR reaction liquid, a digestive enzyme, ligase, a ligase buffer solution, an attachment joint, a fluorescence probe, a Raq enzyme, a qPCR primer and a qPCR reaction liquid, wherein the Raq enzyme, the qPCR primer and the qPCR reaction liquid are used in a qPCR reaction. The combined detection kit can detect mutation sites of BRCA1 / 2 genes of breast cancer at the same time, the experimental result is high in sensitivity, the 5% or below of mutation rate can be detected, the detection sensitivity is greatly improved, the problems in the prior art are overcome, and meanwhile, the whole experiment process meets the requirements for mass and quick detection.

The invention discloses primers, a probe and a kit for detecting the BRCA1 gene expression. The nucleotide sequences of upstream and downstream primers of BRCA1 gene are respectively represented by the SEQ ID No. 1-2. The nucleotide sequence of Taqman fluorescence probe of BRCA1 gene is represented by the SEQ ID No.3. The invention also discloses a detection kit composed of the primers and probe mentioned above, and an application method of the kit. The provided kit can detect samples in different forms such as fresh frozen tissues, paraffin embedded samples, and the like, and has the advantages of high sensitivity, high specificity, and wide application range. The kit comprises premix, a sample can be added directly, a sample does not need to be mixed with enzyme, the operation is convenient and simple, the judgment method becomes more scientific and reasonable, the error generated during the experiment process is reduced, the precision is high, furthermore, the detection speed of the detection kit is quick, and the detection result can be obtained within two hours.

The invention belongs to the field of gene engineering and medical oncology and discloses BRCA1 gene g.41244291delT mutation and its application in breast cancer auxiliary diagnosis. The mutation BRCA1 gene sequence has a g.41244291delT site frame-shift mutation in comparison with BRCA1 normal gene sequence. The invention provides a new breast cancer-causing mutation site, and the mutation site can be used in early diagnosis of breast cancer.

The invention belongs to the fields of gene engineering and tumor medical science, and discloses a mutant gene for breast cancer auxiliary diagnosis and application of the mutant gene. The mutant gene has one or more mutant sites: a mutant site A which has g.41215910delA, g.41244291delT, g.41223130A>T and g.41256139T>A, and other mutation sites which have g.32912699A>G, g.32912799T>C, g.32929399dupT, g.32911757C>T, g.32911659-32911662delAAAA and g.32910963T>G compared with a normal BRCA2 gene sequence. The invention provides a breast cancer pathogenic mutation site combination, and the mutation site combination can be used for early diagnosis of the breast cancer.

A method of predicting the presence of a non-functional BRCA1 gene in a biological sample, comprises the step of assaying the sample for expression of at least one specific member of the S100 family of genes. The invention also describes a kit for predicting the presence of non-functional BRCA1, comprising means for assaying a sample for expression of at least one specific member of the S100 gene family. A method of detecting a genetic predisposition to cancer is also described, comprising the step of assaying a biological sample for expression of a specific member of the S100 family of genes. Also described is a method of determining a suitable chemotherapeutic agent for an individual.

The invention discloses a BRCA1 gene g.43063754T) G mutant and application thereof in breast cancer auxiliary diagnosis, belonging to the field of genetic engineering and tumor medicine. 43063754T) Gsite mutation compare with that normal gene sequence of BRCA1, comprising a specific prim for detecting the mutant, a kit comprising the specific primer, and application of the mutant in breast cancerauxiliary diagnosis. The invention provides a novel mutation site for causing breast cancer, which can be used for early diagnosis of breast cancer.

The invention discloses a primer for simultaneous detection of a BRCA1 / 2 exon sequence and a chemotherapy drug site. The primer comprises: an exon amplification primer pair designed for BRCA1, an exonamplification primer pair designed for BRCA2, and a specific primer pair designed for an SNP site, wherein each exon amplification primer pair is divided into two groups respectively, and no amplification region overlapping exists inside each group of primers of the same gene; in each group of the primer pairs, the 5' end of an upstream primer of a first part primer is added with SEQ ID.NO1, andthe 5' end of a downstream primer is added with SEQ ID.NO2; the 5' end of an upstream primer of a second part primer is added with SEQ ID.NO2, and the 5' end of a downstream primer is added with SEQ ID.NO1; a group of the primer pair of the BRCA1 gene and a group of the primer pair of the BRCA2 gene are randomly combined to form two primer group pairs; and the 5' end of an upstream primer of the specific primer is added with SEQ ID.NO1, and the 5' end of a downstream primer of the specific primer is added with SEQ ID.NO2. The cost is low, throughput is high, a detection range is wide, and a detection time is short; and BRCA1 / 2 and the cancer chemotherapy drug detection site can be simultaneously detected.

The invention discloses a rapid detection kit for breast cancer susceptibility genes, which comprises polymorphic detection of 5 SNP (single nucleotide polymorphism) corresponding to each of Foxp3, FGFR2, CD14 and BRCA1 genes. The invention further discloses a rapid detection method for the breast cancer susceptibility genes. The rapid detection kit and detection method have the benefits as follows: by means of semi-nested AS-PCR (allele-specific polymerase chain reaction) amplification and AGE (agarose gel electrophoresis) detection, genetic typing is performed according to DNA (deoxyribonucleic acid) electrophoretic bands, individuals with potential breast cancer susceptibility are screened out, so that breast cancer is prevented; and the method is simple and convenient to operate, short in detection time, low in cost, high in sensitivity and specificity and broad in application prospect.

The invention discloses a kit for detecting mutation sites of BRCA 1 gene, BRCA 2 gene and PALB 2 gene, comprising ARMS primers shown as SEQ ID NO. 1 to 12 and probes as SEQ ID NO. 20 to 23. In addition, the invention further discloses a method for detecting the mutation sites of the BRCA 1 gene, the BRCA 2 gene and the PALB 2 gene, which comprises the steps of 1) extracting a DNA (deoxyribonucleic acid) sample to be used as a DNA template; 2) performing fluorescent PCR (polymerase chain reaction) amplification on the DNA template by using the ARMS primers shown as the SEQ ID NO. 1 to 12 and the probes shown as SEQ ID NO. 20 to 23; 3) detecting the fluorescence intensity of a reaction system in the fluorescent PCR amplification, and judging the mutation of the mutation sites, namely the 10th exon and the 23th exon of the BRCA 1 gene, the 11th exon of the BRCA 2 gene and the 4th exon of the PALB 2 gene. The kit and the method provided by the invention are capable of overcoming the defects of low flux, long time consumption, limited detecting capability and the like in the existing sequencing technique, and have the advantages of quickness, high flux, good specificity, high sensitivity and the like.

The present invention relates generally to the field of human genetics, and more specifically to the detection of a specific type of germline mutations in the BRCA1 gene, which will predispose to breast and ovarian cancer. In addition, the invention relates to the molecular genetic mechanism that may have mediated the genesis of these mutations, in particular the role of Alu repetitive DNA elements present in the intronic regions of BRCA1. The invention further relates to somatic mutations of this type in the BRCA1 gene in human breast and ovarian cancer, and their use in the diagnosis and prognosis of human breast and ovarian cancer. The invention more particularly relates to the screening of this type of BRCA1 mutations in human genomic DNA, which are useful for the diagnosis of inherited predisposition to breast and ovarian cancer.

The invention relates to a universal BRCA1 gene multi-PCR (polymerase chain reaction) database creating reagent kit, and particularly provides a primer product, a polynucleotide sequence product, a corresponding reagent kit and a BRCA1 gene multi-PCR database creating or BRCA1 gene mutation screening method. Primers in the primer product comprise primer sequences shown as SEQ ID NO:1-84. Each polynucleotide sequence in the polynucleotide sequence product contains suspension linker sequences and an optional primer sequence shown as SEQ ID NO:1-84.

The invention discloses a method of capturing a genomic target sequence for high-throughput sequencing. The method can sequentially include the following steps of (1) preparing a genomic DNA library;(2) hybridizing with a capture probe library, wherein the capture probe library is composed of n specific probes, n is a natural number, and the specific probes are single-stranded DNA molecules whichare designed and synthesized according to the nucleotide sequence of the target sequence and subjected to locked nucleic acid modification. The capture probe library is composed of 440 probes including the LNA probe-1 to the LNA probe-440; the nucleotide sequences of the LNA probe-1 to the LNA probe-440 are sequentially shown as sequence 1 to sequence 440 in a sequence list. Experiments prove that the capture probe library can capture the corresponding target sequences in a BRCA1 gene and a BRCA2 gene. Therefore, the provided method has important application value.

The invention discloses a multiple PCR kit for detecting the eleventh exon mutation of breast cancer susceptibility gene and a preparation method thereof. The PCR amplification primers of the kit are composed of primers pair 1 which consist of SEQ ID NO:1 and SEQ ID NO:2 and primers pair 2 which consist of SEQ ID NO:3 and SEQ ID NO:4. The multiple PCR kit of the invention combines the SSCP technology with the DNA silver staining and can simultaneously screen the mutation conditions of the eleventh exon of BRCA1 gene and the eleventh exon of BRCA2 gene. The invention has the advantages of strong sensitivity, high accuracy, simple and convenient property, etc. and has important significance on the early detection and the forecast of the breast cancer.