52 results about "PIK3CA gene" patented technology

The present invention discloses primers, probes, a detection system and a kit for one time detection of lung cancer multiple genes, wherein the primers, the probes, and the distribution way for detecting 25 EGFR gene mutations, 7 KRAS gene mutations, 6 BRAF gene mutations, 9 NRAS gene mutations, 5 HER2 gene mutations, 2 PIK3CA gene mutations, 5 fusion genes of ALK5, 13 fusion genes of ROS1, and 6 fusion genes of RET are provided. According to the present invention, the detection kit adopts the 12 linking PCR reaction strip design, each 12 linking PCR strip detects multiple genes of a sample, and the corresponding fusion detection reagents and the internal control reagents are filled in the pipes 1-4 of the 12 linking PCR strip; and with the primers, the probes, the detection system and the kit, the one-time detection of the 24 fusions and the 54 mutations of the lung cancer can be achieved, such that the detection time is substantially shortened, the sensitivity is high, the specificity is strong, the operation is simple and rapid, and the reference for selection of tumor targeting drug therapy on lung cancer patients can be provided for clinician.

The invention discloses a liquid phase chip for detecting PIK3CA (phosphoinositide-3-kinase, catalytic, alpha) gene mutation. The liquid phase chip mainly comprises (A) a wild-type and mutant-type allele specific primer extension (ASPE) primer pair designed for mutational sites of a PIK3CA gene respectively, microspheres which are respectively coated with specific anti-tag sequences and have different colors of codes and amplification primers for respectively amplifying the target sequences of the PIK3CA gene with the corresponding mutational sites of the Exon 9 and/or Exon 20, wherein the anti-tag sequences can correspondingly complement and form pair with the selected tag sequences. The liquid phase chip has the following advantages: the coincidence rate of the detection method provided by the invention and a sequencing method is as high as 100%; the prepared liquid phase chip has very good signal to noise ratio; cross reactions do not exist between the designed probes and the anti-tag sequences; and parallel detection of various mutant types and various mutational sites on the same site can be realized.

The invention discloses a kit detecting hot spot mutation sites in a colorectal cancer PIK3CA gene. The kit comprises an erythrocyte lysate, a whole blood DNA extraction reagent, an anhydrous ethanol, a PCR amplification reaction solution, a positive control, a negative control and a pyrosequencing reaction solution. The kit is characterized by comprising upstream and downstream primers 9F-Biotin and 9R for detecting exon 9 of the target gene and a sequencing primer 9RCX, and upstream and downstream primers 20F-Biotin and 20R for detecting exon 20 of the target gene and a sequencing primer 20RCX. The kit can be used to detect polymorphism of hot spot mutation sites (E542K, E545K and H1047R) in the colorectal cancer related PIK3CA gene, and has good specificity and high accuracy.

The invention relates to the field of molecular biology, in particular to a method for detecting gene mutations by adopting fluorescent quantitation PCR, as well as a fluorescent quantitation PCR detection kit and a detection method for PIK3CA (phosphatidylinositol3-kinase catalytic alpha) gene mutations. The method comprises the following steps: designing a specific primer and a probe; preparing a fluorescent quantitation PCR reaction system and designing reaction conditions; amplifying a mutant gene target sequence in a sample by utilizing the specific primer; detecting the fluorescence signal strengths of FAM, HEX and Cy5 in the reaction system, and taking the fluorescence signal strengths as judgement standards of results. The kit provided by the invention can detect 8 types of mutations of a PIK3CA gene simultaneously; the specificity and the selection capability are high; a PIK3CA mutated molecule, which is as low as 5 copy, can be detected in a clinical sample of which the mutation content is only 0.1 percent; the detection speed is high, that is, the detection can be fulfilled within 90 minutes.

The invention discloses a probe, a primer and a kit for detecting drive mutation of a PIK3CA (Phosphatidylinositol-3-kinases) gene and relates to detection of gene mutation. The method disclosed by the invention comprises the steps of: (1) providing 6 groups of primers (18 in number) and the probe; (2) extracting a DNA (deoxyribonucleic acid) template of a sample to be detected; (3) preparing a fluorescent PCR (polymerase chain reaction) system for detecting drive mutation of the PIK3CA gene; and (4) detecting the FAM and HEX fluorescent intensity of the reaction system by using the hybridization of a bi-loop probe and the specific primers, and determining a result according to the FAM and HEX fluorescent intensity. The method disclosed by the invention can detect 5 types of drive mutation of the PIK3CA gene, and has the characteristics of high sensitivity, strong specificity, fast detect speed and the like.

The invention is applicable to biotechnology and the medical field and relates to a reagent kit used for testing PIK3CA (phosphatidylinositol-4,5-bisphosphate3-kinase, catalytic subunit alpha) gene mutations. A first reagent, a second reagent, a third reagent, a fourth reagent and a fifth reagent are sequentially targeted to E542K, E545K, E545D, H1047R and H1047L mutations of a PIK3CA gene, and a sixth reagent is targeted to a conserved region of the PIK3CA gene and used for quality control. A testing method is carried out by the testing reagent kit by means of fluorescence quantitative PCR (polymerase chain reaction). The testing reagent kit is high in sensitivity and good in specificity and is a high-performance human PIK3CA gene mutation testing reagent kit. The fluorescence quantitative PCR method is adopted for performing specific amplification testing on sequences in PIK3CA gene mutation regions, and specific binding of the MGB (minor groove binder) blocked nucleic acid and a wild type DNA (deoxyribo nucleic acid) template in a sample to be tested is utilized to inhibit binding of the wild type DNA template and a primer probe. Forward primers are designed by methods of lowering the primer Tm value, tailing the 5' end and adding and inserting bases among the primers and the like, specificity of the reagent kit can be improved, then more than 45 cycle numbers are introduced, and accordingly effect of improving sensitivity of the regent kit is achieved.

The invention discloses a breast cancer PIK3CA hotspot mutation detection probe and primer sequence combination and a kit thereof. The detection probe and primer sequence combination comprises a primer sequence as shown in SEQ ID NO: 1-4, and a probe sequence as shown in SEQ ID NO: 5-10. The probe with the sequence is modified with a fluorophore. Based on a micro-drop digital PCR platform, primersand probes are designed for three mutation sites of the PIK3CA gene; and through experimental verification, a breast cancer PIK3CA hotspot mutation detection probe and primer sequence combination isdetermined and the combination can effectively detect the three mutation sites of the PIK3CA gene.

The invention discloses a kit for detecting PIK3CA gene mutation and a detecting method thereof. The kit comprises a special probe which is modified by LNA locked nucleic acid and used for PIK3CA gene mutation sites; the special probe can be combined with wild type DNAs, a sample containing 0.01% PIK3CA gene mutation DNAs can be detected, and the minimum detectability ranges from 2 copies to 5 copies. The detecting method has the advantages that as for PIK3CA gene mutation detection, the specificity is high, the flexibility is high, pollution is small, operation is easy and rapid, and the safety performance is high; the detecting method is particularly suitable for detecting gene mutation from body fluid such as plasma, urine and saliva with low mutation content, and is suitable for early screening and diagnosis of colorectal cancers, and guidance is provided for personalized medicines.

The present disclosure relates to methods of treating a vascular malformation in a subject expressing a gain-of-function mutation in a PIK3CA gene comprising administering, to the subject, an effective amount of an agent that inhibits phosphoinositide 3-kinase (“PI3K”).

The invention relates to a kit for detecting effectiveness of oxaliplatin to colorectal cancer. The kit comprises a pair of amplification primer sequences for detecting MAP4K1 gene copy number, a pair of amplification primer sequences for detecting PIK3CA gene copy number and a pair of primer sequences for detecting reference gene GAPDH gene copy number. Through the kit designed in the invention, effectiveness of oxaliplatin to colorectal cancer can be detected rapidly and effectively, and whether a patient to be detected is suitable to use oxaliplatin as a first-line chemotherapy drug is evaluated. The kit provided by the invention provides effective reference for clinical medication.

The invention discloses a kit for detecting 34 mutation sites of lung cancer based on MALDI-TOF-MS. The kit comprises multiple PCR amplification reaction primers which are shown as SEQ ID NO:1-16, andsingle-base extension primers which are shown as SEQ ID NO:17-48. The kit provided by the invention can be used for simultaneously detecting 23 sites of an EGFR gene, 7 sites of a KRAS gene, 1 site of a BRAF gene and 3 sites of a PIK3CA gene. According to the kit provided by the invention, the 34 sites of the four genes can undergo target segment amplification in two holes and single-base extension in eight holes; and with the application of the kit provided by the invention, 34 site mutations of the four genes, which may exist in an abnormal sample, can be simultaneously detected and analyzed. The kit provided by the invention has important guiding significance for mastering related factors of a patient on drug susceptibility, conducting individualized treatment on the patient with the lung cancer and targeting to drug adaptability.

A polynucleotide comprising at least the final six nucleotides of one of the following primer sequences, or a sequence complementary thereto: SEQ. ID NOS. 3 to 16, 18, 20 to 33, 35 or 37 to 39. A method of detecting the presence or absence of a mutation in the PIK3CA gene, wherein the mutation is one of H1047R, H1047L, E542K and E545K, and preferably ARMS primers are combined with Scorpion primers.

Disclosed is a kit for detecting drug resistance of cetuximab in the treatment of metastatic colorectal cancer. The kit comprises a substance used for detecting gene mutations in Exon 19 of the PIK3CA gene, and may further comprise a specification recording the following contents: if Exon 19 in the PIK3CA gene of a patient with metastatic colorectal cancer as a subject to be tested, who is intended to receive cetuximab treatment or is receiving cetuximab treatment and does not have drug resistance, has at least one of K944N, F930S, V955G, V955I, and K966E mutations, the subject to be tested will develop drug resistance or will be a candidate to develop drug resistance when receiving or continuing to receive cetuximab for treating metastatic colorectal cancer.

The invention relates to the field of biotechnology, and provides a primer and a method for mass spectrometric detection of hotspot mutation of PIK3CA genes by using the primer. The method comprises the following steps of: (1) positioning hotspot mutation sites of the PIK3CA genes; (2) designing a primer for amplified reaction and a primer for extension reaction according to deoxyribonucleic acid (DNA) sequences at two ends of the mutation sites; (3) performing the amplified reaction; (4) performing treatment by using a serum amyloid protein (SAP) enzyme (alkaline phosphatase); (5) performing the extension reaction; (6) purifying a product of the extension reaction by using resin; (7) transferring the product purified by the resin to a chip; (8) putting the chip in a mass spectrograph, and running a detection procedure; and (9) reading and analyzing detection data, and determining the gene type of a target site of the PIK3CA genes. Compared with the conventional detection method, the method provided by the invention has the characteristics of high sensitivity, high accuracy, high flux and low cost.

The invention discloses a laying duck follicle growth-associated miRNA alpha-mir-145 and detecting primer, inhibitor and application thereof and belongs to the technical field of genetic engineering.The nucleotide sequence of the laying duck follicle growth-associated miRNA alpha-mir-145 is shown as SED ID NO: 1. Research shows that the laying duck follicle growth-associated miRNA alpha-mir-145 can regulate follicle growth through targeted PIK3CA genes, reduce expression of a cell proliferation marker gene CyclinB2 and enhance expression of a cell apoptosis marker gene BCL2. The laying duck follicle growth-associated miRNA alpha-mir-145 can serve as a marker for detecting proliferation of laying duck follicular cells, thereby filling the gap in the field, providing a theoretical basis anda scientific basis for analysis on the genetic mechanism of laying duck follicle growth and being of great significance to researching the genetic nature of duck reproduction traits, improving the reproductivity of laying ducks and achieving assisted genetic breeding.

The invention provides a primer, detection method and kit for detecting human PIK3CA gene E545K mutation. The detection primer provided by the invention can be combined with a specific sequence of a PIK3CA gene E545K to amplify a mutation sequence. The detection method provided by the invention can be combined with an amplified fragment by adopting a fluorescence signal detection group to send outa fluorescence signal; a blocking agent is specifically combined with a wild type sequence corresponding to a mutation site to inhibit wild type non-specific amplification. A competitive allele specific PCR (Polymerase Chain Reaction) amplification technology is adopted to establish the kit based on real-time fluorescence PCR and the PIK3CA gene E545K mutation can be accurately detected. The method provided by the invention has the advantages of simplicity in operation, easiness of reading results and high sensitivity, and can be used for detecting a sample containing 0.001 percent of the PIK3CA gene E545K mutation; ultrasensitive detection of the PIK3CA gene E545K mutation is realized.

The invention provides a primer-probe composition for detecting a PIK3CA gene at the E545K site and a detecting method thereof. The primer-probe composition comprises specific primers SEQ ID NO.1-2 aiming at the PIK3CA gene at the E545K site, a mutant probe SEQ ID NO.3 aiming at the PIK3CA gene at the E545K site and a wild probe SEQ ID NO.4 aiming at the PIK3CA gene at the E545K site, by designingthe primers and modifying the probes aiming at the specific area of the E545K site, the primers and the probes are matched with one another, ddPCR and the primer-probe composition with the high specificity are combined to optimize extraction of cfDNA, recurring number and annealing temperature of PCR, each step synergizes and interacts to improve the accuracy, the stability and the sensitivity ofdetection ultimately, and the primer-probe composition and the detecting method thereof have wide application prospects and market value.

The invention relates to detection of a mutation site of a PIK3CA gene in the ctDNA in urine, and particularly provides a method for separating the ctDNA from a urine sample, a method for building a sequencing library based on the urine sample, a method for performing nucleic acid sequencing based on the urine sample according to the sequencing library, equipment for separating the ctDNA from theurine sample and a system for determining the sensitivity of an individual drug. The method for separating the ctDNA from the urine sample comprises the following steps: adding EDTA into the urine sample to obtain a first mixed solution; centrifugating the mixed solution, and collecting supernate; concentrating the supernate to obtain a concentrated solution; mixing the concentrated solution withanion exchange resin to obtain a second mixed solution; performing chromatographic separation treatment on the second mixed solution to obtain eluent; purifying the eluent to obtain the ctDNA. According to the detection disclosed by the invention, a large quantity of ctDNAs can be extracted quickly from a large quantity of urine samples, and can be subjected to nucleic acid sequencing, and the sensitivity of the individual drug can be determined, so that the application prospect of the ctDNA in the urine is enlarged.