34 results about "Breast cancer susceptibility genes" patented technology

The invention discloses a hybrid capture kit for detecting mutation of breast cancer susceptibility genes BRCA1 and BRCA2. The hybrid capture kit comprises a hybridization reagent, a PCR amplification reagent and an enrichment degree detection reagent, wherein the hybridization reagent comprises probe mixtures from SEQ NO.1 to SEQ NO. 173, and the mole ratio is SEQ NO.1: SEQ NO. (2-172): SEQ NO. 173=1: 1: 1. During detection, the use amount of each of the probe mixtures with the concentration of 2nM is 1 [mu]l. The kit disclosed by the invention can be used for high-sensitivity, high-flux, low-cost and easy-operation full-explicit mutation detection for the genes BRCA1 and BRCA2, and can assist a clinical doctor in screening out individuals with the breast cancer susceptibility risk to fulfill the aim of preventing occurrence of breast cancers.

The invention discloses a primer set, a method and a kit for Long-range PCR (polymerase chain reaction) detection of a BRCA (breast cancer susceptibility gene) 1 and a BRCA 2. The primer set comprises 18 primer pairs altogether, wherein the primer pair 1 comprises a forward primer BR1_P6.1F: 5'-ACGTTGCGGAGAGGTGAG-3' and a reverse primer BR1_P6.1R: 5'-TAGCCAGGCATAGTTGCACA-3'; the second primer pair comprises a forward primer BR1_P6.2F:5'-CTCCACCTCCCTGGTTCAGT-3' and a reverse primer BR1_P6.2R:5'-AGCTTTGTGGTGAGGTGTTG-3'; the third primer pair comprises a forward primer BR1_P7F:5'-GGCTCAAAGGACCTCCTACC-3' and the reverse primer BR1_P7R:5'-AGGCAGAGGAACCTCTT GAA-3'. The primer set, the method and the kit have the benefits that the detection method requires few primers, enrichment can be effectively performed on BRCA1 and BRCA2 target areas, and single amplified bands are clear. The time is obviously shortened with the technique, and costs are also greatly reduced.

The invention discloses a detection analysis method for a breast cancer susceptibility gene heritable variation point. The detection analysis method is characterized by comprising the following steps: (1) a data quality control step; (2) a sequence alignment step; (3) variation detection step; (4) a variation annotation step; and (5) a statistical report step. The detection analysis method has the advantages of integration, high efficiency and visualization.

The invention relates to the field of genetic engineering, and provides a method and a kit for assaying breast cancer susceptibility genes. The method for assaying breast cancer susceptibility genes includes the following steps: (A) breast cancer susceptibility gene specific primers are utilized to amplify a plurality of target regions in a sample to be assayed, and a sequencing library is constructed on the basis of amplification products; (B) the single-molecule amplification of the sequencing library is carried out, so that a plurality of single-molecule amplification products corresponding to the target regions are obtained; (C) the high-throughput gene sequencing of the single-molecule amplification products is carried out simultaneously, so that the sequence information of the target regions is obtained. The method and the kit for assaying breast cancer, which are provided by the invention, can assay a plurality of regions of a breast cancer susceptibility gene at the same time, consequently, the efficiency of assay is increased, the assay cost is reduced, and in addition, the sensitivity of assay and the comprehensiveness and accuracy of the assay result are increased as well.

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 mammary-cancer-susceptible-gene detection chip which comprises a solid support and a gene detection probe fixed onto the solid support. The gene detection probe comprises 31 SNP (single-nucleotide polymorphism) loci of 24 mammary-cancer-susceptible genes. The 24 mammary-cancer-susceptible genes are respectively CHEK2, PALB2, ZNF350, ESR1, C6orf97, USP7, MRPS30, AURKA, ATM, CASC16, TOX3, DNMT1, ZMIZ1, TGFB1, FGFR2, ZNF365, SLC4A7, TNRC9, TNP1, HER2, BRCA2, CCDC170, BRCA1 and CYP11A1. The gene detection probe comprises the 31 SNP loci of the 24 mammary-cancer-susceptible genes. The documents prove that the loci have higher practicality, can be used for large-scale screening of early diagnosis of diseases, have the advantages of high detection success rate, favorable technique reproducibility and high cost performance, and provide important references for diagnosis, typing and prognosis of mammary cancer, thereby implementing early diagnosis of diseases.

The invention discloses a breast cancer susceptibility gene BRCA1 and BRCA2 detection kit and method. The kit includes a first-round PCR reagent and a second-round PCR reagent. The first-round PCR reagent contains a buffer solution, magnesium salt, deoxyribonucleoside triphosphate, enzyme mixture liquid and a first-round PCR primer. The second-round PCR reagent contains a buffer solution, magnesium salt, deoxyribonucleoside triphosphate, enzyme mixture liquid and a second-round PCR primer. The first-round PCR primer includes 46 pairs of primers SEQ1-SEQ92 and 67 pairs of primers SEQ93-SEQ226, the 5'end of each upstream primer is added with a sequence SEQ227, and the 5'end of each upstream primer is added with a sequence SEQ228. The primer combination includes 113 pairs of primers and covers all the coding sequences and adjacent at least +5bp intron subdomains of a breast cancer susceptibility genes. An NGS technology is adopted to detect mutations of all the coding sequences and splice sites of the breast cancer susceptibility genes BRCA1 and BRCA2, the kit has very high specificity and accuracy, and the method is simple, convenient and efficient.

The invention discloses a breast cancer susceptibility gene BRCA2 site g.32336534T) C mutant and application thereof, belonging to the field of pharmaceutical biology. By PCR- A specific primer technique is used to design a primer sequence that can amplify a region containing a target site of a mutated gene sequence. The invention provides a novel mutation site for causing breast cancer, which canbe used for early diagnosis of breast cancer.

The invention discloses a human BRCA1 / BRCA2 gene mutation detection kit, which comprises an amplification box for amplifying all exons of breast cancer susceptibility genes (BRCA1 / BRCA2) and at least10bp of DNA sequences at the upstream and downstream of each exon, an amplification product library building box, a library building connection specificity connector box, a detection kit outer package box and a magnetic bead box. The amplicon capture sequencing technology is used; the difficulty and the complexity of the conventional probe capture sequencing technology are greatly reduced; meanwhile, the technical cost is effectively reduced; the popularization and the use of clinical hereditary breast cancer and ovarian cancer screening detection are facilitated.

The invention belongs to the field of gene detection, and in particular relates to a method for detecting genes related to early diagnosis of breast cancer based on nucleic acid mass spectrometry technology. The present invention provides a combined detection method for 15 breast cancer-related mutation sites of 4 genes and 18 breast cancer susceptibility SNP sites of 10 genes, and uses SequenomMassARRAYSNP genotype analysis technology to detect SNPs of breast cancer susceptibility genes Genotyping detection of loci and gene mutation loci associated with breast cancer. This method is more accurate for early screening of breast cancer, has good technical reproducibility, and is cost-effective; compared with TaqMan-PCR and SNP detection of second-generation platform sequencing, it has flexible design, high accuracy, and large sample size of detection sites , low cost and other advantages, hundreds to thousands of samples can be tested on dozens of SNP sites at the same time.

The present invention relates to a rapid sensitive breast cancer susceptibility gene SNP detection method based on PCR primer 3'terminal nucleotide dideoxy modification. Aiming at breast cancer susceptibility gene single nucleotide polymorphism (SNP), the invention discloses a novel detection method based on primer 3' terminal dideoxy modification, the flexible, convenient and strong operability features of primer extension method are retained, the production of false positive is fundamentally solved, and a rapid, economical and accurate sensitive cancer susceptibility gene and other disease-related gene SNP detection method is provided.

The invention relates to a breast cancer susceptibility gene variable library construction method. The construction method comprises the following steps that 1, according to variation regions of a breast cancer-related gene, primer pairs capable of detecting the variation regions are designed; 2, judgment parameters R of the primer pairs are calculated, the primer pairs of which values of the judgment parameters are smaller than or equal to 1 are classified into a first primer combination solution, and the primer pairs of which values of the judgment parameters are larger than 1 are classified into a second primer combination solution; 3, a to-be-detected DNA sample is extracted and purified; 4, initial PCR amplification is conducted on the purified sample by adopting the first primer combination solution and the second primer combination solution separately; 5, linker connection is conducted on a target fragment to obtain a fragment with a linker; 6, library PCR amplification is conducted through a mixed solution of the first primer combination solution and the second primer combination solution to obtain a sequencing library. The library constructed through the breast cancer susceptibility gene variable library construction method is high in sequencing flux, sensitivity and specificity and can be used for detecting low-frequency mutation of free DNA.

The invention provides nucleotide sequences for amplification testing of HER-2 gene copy number, a kit and application thereof. The nucleotide sequences contain 8 pairs of PCR amplification primers for amplification of BRCA1 (Gene ID: 672) full-length gene and 8 pairs PCR amplification primers for amplification of BRCA2 (Gene ID: 675) full-length gene. DNA trapped by amplification can be used forsubsequent high-throughput sequencing analysis. In comparison with a probe targeting trapping technology and a multiplex PCR technology, the method of the invention is simple to operate, is low-cost,has good sequencing data uniformity, can reach more than 99% coverage with small sequencing data amount, and is helpful for providing gene detection technical support for risk assessment or medicationof genetic tumors related with breast cancer susceptibility genes and establishment of a database.

The invention provides a construction method of a BRCA (breast cancer susceptibility gene) high-throughput sequencing library and belongs to the field of high-throughput sequencing. The construction method provided herein comprises generating an initial library, purifying the initial library, generating a sequencing library and purifying the sequencing library. The whole construction method provided herein is performed in a vessel, so that cross infection probabilities are avoided. The construction method provided herein allows purification to be efficient and convenient since magnetic bead reaction is combined with washing operation. The construction method provided herein allows utilization rate of a template to be increased greatly.

The invention discloses a screening method for colorectal cancer susceptibility genes, which is quick and accurate and can cover all exon areas of newest colorectal cancer susceptibility genes. According to a basic scheme of the invention, the screening method for the colorectal cancer susceptibility genes comprises the following steps: extracting 3 to 5 ml of blood from an individual; extracting3 to 5 mu g of gDNA from the blood; breaking and amplifying the gDNA by utilizing ultrasound so as to construct a whole genome library of human, then, capturing breast cancer susceptibility genes by utilizing a colorectal cancer susceptibility gene scanning kit disclosed by the invention, and later on, performing high-throughput sequencing by utilizing a new generation sequencing instrument, and analyzing mutation information related to the genes to achieve the purpose of screening the colorectal cancer susceptibility genes.

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 provides a breast cancer susceptibility gene detection reagent kit. The breast cancer susceptibility gene detection reagent kit comprises 12 primer groups for carrying out PCR (polymerase chain reaction) amplification on mutation hotspots of breast cancer susceptibility genes BRCA1 and BRCA2. The breast cancer susceptibility gene detection reagent kit has the advantages that the breast cancer susceptibility genes are basically identical in PCR amplification efficiency, accordingly, PCR amplification products can be used for second-generation sequencing on DNA (deoxyribonucleic acid) fragments with sequencing requirements, and ultimately obtained sequencing data for the mutation hotspots are excellent in uniformity.

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.

It has been discovered that the human GT198 gene (gene symbol PSMC3IP) at chromosome 17q21 acts as a tumor suppressor. The mutation of the GT198 gene causes the increased dominant negative splice variant activity and leads to the loss of wild type GT198 function, and in turn, induces breast and ovarian cancers. One embodiment provides compositions and methods for treating or alleviating one or more symptoms associated with cancer due to the GT198 gene mutations. Another embodiment provides methods and compositions for detecting cancer due to the mutation of the GT198 gene. Still another embodiment provides methods for identifying compounds, antibodies and natural product molecules that are useful for treating cancer due to the mutations of the GT198 gene. Preferably the disclosed compositions antagonize or interfere with the biological activity of splice variants of GT198.

The invention discloses an LSP (linear scorpion primer) and kit for detecting human BRCA1 (breast cancer susceptibility gene 1) mutations, wherein 5' end of the LSP is provided with a linear probe having different dual-fluorescent labels, 3' end is provided with a sequence fragment complementary to a template, and the two sequence fragments are linked via spacer 18; when the template exists, the LSP and the template can be extended by annealing to synthesize a template complementary chain; after the template complementary chain is synthesized, the 5' end probe portion of the LSP may bind with the chain, forming a scorpion-like shape; a downstream primer is extended along the 3' end of the template complementary chain; the probe portion is sheared under the action of a polymerase so as to emit light. The LSP of the invention has good specificity and high sensitivity, approximate genes are free of mutual interference, and detection results show higher fluorescent signal-to-noise ratio.

The invention relates to a breast cancer susceptibility gene variable library construction method. The construction method comprises the following steps that 1, according to variation regions of a breast cancer-related gene, primer pairs capable of detecting the variation regions are designed; 2, judgment parameters R of the primer pairs are calculated, the primer pairs of which values of the judgment parameters are smaller than or equal to 1 are classified into a first primer combination solution, and the primer pairs of which values of the judgment parameters are larger than 1 are classified into a second primer combination solution; 3, a to-be-detected DNA sample is extracted and purified; 4, initial PCR amplification is conducted on the purified sample by adopting the first primer combination solution and the second primer combination solution separately; 5, linker connection is conducted on a target fragment to obtain a fragment with a linker; 6, library PCR amplification is conducted through a mixed solution of the first primer combination solution and the second primer combination solution to obtain a sequencing library. The library constructed through the breast cancer susceptibility gene variable library construction method is high in sequencing flux, sensitivity and specificity and can be used for detecting low-frequency mutation of free DNA.

The invention discloses a breast cancer susceptibility gene BRCA2-K2305N mutant and a specific primer thereof, and belongs to the technical field of molecular biology. A primer sequence containing a changed mutant gene is designed by applying a PCR-specific primer technology, and for a specific BRCA2 gene g.32344631G>Tmutation site is subjected to PCR amplification detection. The invention provides a new mutation site of a breast cancer related gene, which can be used for early breast cancer screening.

The invention discloses a primer pair, a kit and a method for detecting breast cancer susceptibility gene variation based on long fragment PCR (Polymerase Chain Reaction). The specific breast cancer susceptibility gene variation long fragment PCR detection primer pair is designed, PCR reaction and gene sequencing are combined, rapid and low-cost detection of breast cancer susceptibility gene variation is achieved, the detection rate reaches up to 100%, the specificity is 100%, and the method can be used for clinical detection of breast cancer susceptibility gene variation.

The invention relates to a PCR primer and application of the coding sequences of human breast cancer susceptibility genes BRCA1 and BRCA2 based on NGS technology. The PCR primers of the human breast cancer susceptibility genes BRCA1 and BRCA2 coding sequences include at least one pair of capture primers, and the sequences of the forward primer and reverse primer of each pair of capture primers include specific sequences and are compatible with the described Linker sequence to which the 5' end of the specific sequence is ligated. The PCR primers and applications for amplifying the coding sequences of human breast cancer susceptibility genes BRCA1 and BRCA2 based on NGS technology, the experimental operation involved in the whole method is simple, only involves PCR reagents and primer combinations, the cost is low, and double tags are introduced at the same time Sequence adapter sequence is used to distinguish different samples, which can realize high-throughput sample sequencing detection, and can effectively provide genetic detection support for risk assessment of human breast cancer and other BRCA susceptibility genes-related hereditary tumors or targeted drugs for BRCA mutations .

The invention belongs to the field of breast cancer screening, and particularly relates to a breast cancer susceptibility gene screening method. Aiming at the problems that only breast cancer susceptibility gene screening can be carried out and screened patient data cannot be collected and classified in the prior art, the invention provides the following scheme that the method comprises the following steps: S1, extracting 5ml of venous blood, extracting genome DNA, and carrying out bioinformatics detection; S2, if the detection is negative, determining that the group is a breast cancer low-risk group, and if the detection is positive, determining that the breast cancer risk is very high; S3, printing a detection report, and performing keyword extraction on the detection report; S4, if negative words are extracted, automatically rejecting the extracted keywords; S5, if the positive characters are extracted, extracting the patient information of the detection report; S6, collecting and classifying the extracted information. The information can be collected and classified, a patient can conveniently refer to the information, and preliminary self-examination is completed.

The invention relates to the field of genetic engineering, and provides a method and a kit for assaying breast cancer susceptibility genes. The method for assaying breast cancer susceptibility genes includes the following steps: (A) breast cancer susceptibility gene specific primers are utilized to amplify a plurality of target regions in a sample to be assayed, and a sequencing library is constructed on the basis of amplification products; (B) the single-molecule amplification of the sequencing library is carried out, so that a plurality of single-molecule amplification products corresponding to the target regions are obtained; (C) the high-throughput gene sequencing of the single-molecule amplification products is carried out simultaneously, so that the sequence information of the target regions is obtained. The method and the kit for assaying breast cancer, which are provided by the invention, can assay a plurality of regions of a breast cancer susceptibility gene at the same time, consequently, the efficiency of assay is increased, the assay cost is reduced, and in addition, the sensitivity of assay and the comprehensiveness and accuracy of the assay result are increased as well.

The invention belongs to the technical field of breast cancer susceptibility gene screening, and particularly relates to a breast cancer susceptibility gene screening method. Aiming at solving the problems that existing breast cancer susceptibility gene screening has no proper technical means and cannot effectively nip in the bud, the invention provides the following scheme. The method comprises the following steps: S1, creating a gene bank; S2, extracting a comparison sample; S3, the patient is inquired and recorded; S4, analyzing and comparing the sample genes; S5, judging the possibility that the sample gene infects the breast cancer; S6, performing prevention prompting and treatment methods according to the judgment; and S7, generating a report. According to the method, the genes are screened in a comparison mode, firstly, analysis is carried out in two comparison modes, and the similarity of the susceptible genes can be judged; and through secondary classification and comparison of symptoms of patients, the similarity of the susceptible genes can be further calibrated, and the screening effect and efficiency are improved.

It has been discovered that the human GT198 gene (gene symbol PSMC3IP) at chromosome 17q21 acts as a tumor suppressor. The mutation of the GT198 gene causes the increased dominant negative splice variant activity and leads to the loss of wild type GT198 function, and in turn, induces breast and ovarian cancers. One embodiment provides compositions and methods for treating or alleviating one or more symptoms associated with cancer due to the GT198 gene mutations. Another embodiment provides methods and compositions for detecting cancer due to the mutation of the GT198 gene. Still another embodiment provides methods for identifying compounds, antibodies and natural product molecules that are useful for treating cancer due to the mutations of the GT198 gene. Preferably the disclosed compositions antagonize or interfere with the biological activity of splice variants of GT198.

The invention discloses an LSP (linear scorpion primer) and kit for detecting human BRCA1 (breast cancer susceptibility gene 1) mutations, wherein 5' end of the LSP is provided with a linear probe having different dual-fluorescent labels, 3' end is provided with a sequence fragment complementary to a template, and the two sequence fragments are linked via spacer 18; when the template exists, the LSP and the template can be extended by annealing to synthesize a template complementary chain; after the template complementary chain is synthesized, the 5' end probe portion of the LSP may bind with the chain, forming a scorpion-like shape; a downstream primer is extended along the 3' end of the template complementary chain; the probe portion is sheared under the action of a polymerase so as to emit light. The LSP of the invention has good specificity and high sensitivity, approximate genes are free of mutual interference, and detection results show higher fluorescent signal-to-noise ratio.

The invention discloses a rapid detection kit for breast cancer susceptibility genes and a detection method thereof. The breast cancer susceptibility genes refer to SNP sites of CYP1B1 genes, the rapid detection kit detects the SNP sites of the CYP1B1 genes, the kit includes specific amplification primers of the SNP sites of the CYP1B1 genes, Taq enzyme, dNTs and a buffer solution. The SNP sites of the CYP1B1 genes include rs1056827 and rs1056836. A multiplex PCR amplification method is adopted, meanwhile the SNP sites of the breast cancer susceptibility genes are amplified, the amplificationefficiency is greatly improved, and the amplification cost is reduced. A detection result of the kit can be used for pre-judgment, initial check and other aspects of incidence of the breast cancer clinically and can be used for medication guidance and a prognostic test of the breast cancer, after statistics, the detection result can serve as a regional basis for breast cancer research and evaluation, and data support is provided for further revealing the pathogenesis of the breast cancer. The rapid detection kit can be applied to other SNP site detection and has good popularization value.