30 results about "PIK3CA Gene Mutation" patented technology

The invention discloses a PIK3CA gene mutation site detection probe, a detection liquid phase chip and a detection method thereof. The PIK3CA gene mutation detection liquid phase chip mainly comprises a microball covered with a probe and a primer for amplifying a target sequence carrying a 9th exon and / or a 20th exon. The PIK3CA gene mutation detection liquid phase chip and the detection method can help simultaneously detect the sites with relatively higher frequency of the PIK3CA gene mutation, and can separately detect the 9th exon and the 20th exon or detect the 9th exon and the 20th exon at the same time, and the detection has uniform reaction condition, good specificity of detection results, high sensitivity, good accuracy up to 90% and short detection time.

The present invention provides a liquid chip for detecting PIK3CA gene mutations, which comprises wild-type and mutant ASPE primer pairs designed according to the mutational sites of PIK3CA gene; beads coded by different colors and coated with specific anti-tag sequences, wherein said anti-tag sequences are complementary with the tag sequences selected; and amplification primers for amplifying target sequences of PIK3CA gene which comprise the corresponding mutational sites of Exon 9 or Exon 20. The present invention also provides specific primers for detecting PIK3CA gene mutations.

The invention discloses a PIK3CA gene mutation detection system and a kit thereof. The PIK3CA gene mutation detection system comprises primers, a probe and an amplification blocking primer, wherein the primers include a general forward primer, detecting reverse primers and a quality-control reverse primer. The detecting reverse primers include the detecting reverse primer for the E542K mutation site of a PIK3CA gene, the detecting reverse primer for the E545K mutation site of the PIK3CA gene, the detecting reverse primer for the H1047R mutation site of the PIK3CA gene and the detecting reverse primer for the H1047L mutation site of the PIK3CA gene. The 5' end of a probe sequence is provided with a fluorescent marker, and the 3' end of the nucleotide sequence of the probe is provided with another fluorescent marker. The 3' end of the nucleotide sequence of the amplification blocking primer is provided with a phosphorylated modifier. The PIK3CA gene mutation detection system and the kit of the system can detect the E542K, E545K, H1047R or H1047L mutation site of the PIK3CA gene and are high in sensitivity, strong in specificity and fast and convenient to use.

The invention discloses a kit utilizing stem loop primers to detect PIK3CA gene mutation sites. The kit utilizing the stem loop primers to detect the PIK3CA gene mutation sites comprises a set of stemloop primers, and two MGC fluorescent probes; sequence of upstream primers of the set of the stem loop primers is shown as SEQ ID NO. 1 while sequence of downstream primers of the set of the stem loop primers is shown as SEQ ID NO. 8; and the two MGC fluorescent probes are shown as SEQ ID NO. 9-10. The kit utilizing the stem loop primers to detect the PIK3CA gene mutation sites is capable of overcoming defect of low throughput of existing first-generation sequencing technologies, long time consumption of existing second-generation sequencing technologies, limited detection capacity of fluorescent quantitative PCR and the like; moreover, the kit has the advantages of being high in sensitivity, good in specificity, rapid, high in throughput and so on.

The invention discloses a PIK3CA gene mutation site detection probe, a detection liquid phase chip and a detection method thereof. The PIK3CA gene mutation detection liquid phase chip mainly comprises a microball covered with a probe and a primer for amplifying a target sequence carrying a 9th exon and / or a 20th exon. The PIK3CA gene mutation detection liquid phase chip and the detection method can help simultaneously detect the sites with relatively higher frequency of the PIK3CA gene mutation, and can separately detect the 9th exon and the 20th exon or detect the 9th exon and the 20th exon at the same time, and the detection has uniform reaction condition, good specificity of detection results, high sensitivity, good accuracy up to 90% and short detection time.

The invention discloses primers and probes and a kit for detecting five mutation types of a human PIK3CA gene. According to each group of the five groups (PIM1-PIM5) of the primers and probes, the mutation primer can be combined with a conserved sequence of the PIK3CA gene, a mutation ARMS primer can be specially combined with a corresponding mutation sequence of the mutation ARMS primer to amplify the mutation sequence, the detection probe can be combined with an amplified fragment, and the blocking probe can be specially combined with a wild type sequence corresponding to a mutation locus to inhibit wild type nonspecific amplification. Accordingly, by adopting a specific mutation primer and probe blocking technology, the five mutation types of the PIK3CA gene can be accurately detected; through the established kit of a real-time fluorescence PCR amplification reaction system, rapid detection on the PIK3CA gene mutation is facilitated, operation is easy, and a result is easy to read; the detection method is high in sensitivity, and 50-copy mutation can be stably detected; the specificity is good, no nonspecific amplification is generated in 20-ng wild type genome DNA, and the detectability reaches up to 1%.

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.