108results about How to "Precise Quantitative Analysis Features" patented technology

The invention discloses a specific primer, a liquid-phase chip and a method for SNP detection of CYP2C9 and VKORC1 genes. The liquid-phase chip comprises wild-type and mutable-type ASPE primer pairs and microspheres coated by a specific anti-tag sequence respectively, which are designed respectively aiming at each type of mutable loci, primers used for amplifying a CYP2C9 gene target sequence having CYP2C9*2, CYP2C9*3, CYP2C9*5 and CYP2C9*6SNP loci, and / or primers used for amplifying a VKORC1 gene target sequence having G1639A, G1173T and G3730A SNP loci. The liquid phase chip of the invention has a quite good signal-noise ratio, and the cross reaction does not happen between a designed probe and the anti-tag sequence basically; the ASPE primer designed by the invention has quite good specificity, and can accurately differentiate various types of mutable loci; and the detection method has the advantages that: a few steps are adopted, 7 types of SNP loci can be detected in one step, the operation is convenient, a lot of uncertain factors existing in a process of repeated operations can be avoided, and the detection accuracy is greatly improved.

The invention provides a CYP2D6 gene mutation detection liquid-phase chip which comprises ASPE (Allele Specific Primer Extension) primers aiming at CYP2D6 C2850T and CYP2D6Deletion mutational sites, three microballoons respectively enveloped with a specific anti-tag sequence and amplification primers aiming at the CYP2D6C2850T and the CYP2D6 Deletion mutational sites. The CYP2D6 gene mutation detection liquid-phase chip can simultaneously detect aiming at the CYP2D6 C2850T and the CYP2D Deletion mutational sites and has excellent signal to noise ratio. The coincidence ratio with a sequencing method of the CYP2D6 gene mutation detection liquid-phase chip reaches up to 100 percent, and the CYP2D6 gene mutation detection liquid-phase chip has higher specificity and precision compared with intra-class correlation products.

The invention discloses an NRAS gene mutation detection specificity primer and a liquid chip thereof. The liquid chip mainly comprises: an ASPE primer composed of a 5'-terminal tag sequence and 3'-terminal specificity primer sequences focused on target gene mutation sites, wherein the specificity primer sequences comprise SEQ ID NO.18, SEQ ID NO.19, SEQ ID NO.20, SEQ ID NO.21 and / or SEQ ID NO.22 focused on a Codon12 site, SEQ ID NO.23, SEQ ID NO.24, SEQ ID NO.25, SEQ ID NO.26 and / or SEQ ID NO.27 focused on a Codon13 site, SEQ ID NO.28 and SEQ ID NO.29 focused on a Codon18 site, and / or SEQ ID NO.30, SEQ ID NO.31, SEQ ID NO.32, SEQ ID NO.33 and / or SEQ ID NO.34 focused on a Codon61 site; a microsphere coated by an anti-tag sequence; and an amplimer. The consistency between the detection result of the detection liquid chip provided by the invention and the detection result of a sequencing method is high to 100%, and the wild-type and mutant parallel detection of a plurality of mutation sites is realized.

The invention discloses a liquid phase chip for detecting EGFR (epidermal growth factor receptor) gene mutation. The liquid phase chip mainly comprises a wild-type and mutant-type allele specific primer extension (ASPE) primer pair designed for mutational sites of an EGFR 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 with the corresponding mutational sites of the Exon 19, Exon 20 and / or Exon 21, wherein each ASPE primer comprises the tag sequences at the 5' terminal and the specific primers aiming at the mutational sites of the target gene at the 3' terminal; and the anti-tag sequences can correspondingly complement and form pair with the tag sequences selected in (A). 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; and cross reactions do not exist between the designed probes and the anti-tag sequences.

The invention relates to a PCR primer involving 47 genes and a tag PCR primer. The tag PCR primer is composed of the PCR primer and a DNA tag sequence connected to the 5' end of at least one primer in the PCR primer pair, wherein the DNA tag sequence is selected from SEQ ID NO: 107 to SEQ ID NO: 126. The invention further relates to a method and a kit for constructing a sequencing library by applying the DNA tag of the tag PCR primer. The coincidence rates of the detection method provided by the invention and the sequencing method is up to 100%, the provided amplification primer and the DNA tag form a detection product with a specificity, a sensitivity and a repeatability which are optimized and balanced, somatic mutation and mononucleotide polymorphism can be detected in parallel, and a plurality of base sequences of 1-20 sample sources can be accurately differentiated by one-time detection.

The invention discloses an SNP (Single Nucleotide Polymorphism) detection liquid phase chip for KIF6 gene, mainly comprising an ASPE (Application Solid Phase Extraction) primer pair, microballons and an amplification primer, wherein each ASPE primer consists of a 5'-end tag sequence and 3'-end specific primers aiming at T2155C SNP locus; the 3'-end specific primers comprise SEQID NO.23 and SEQ IDNO.24; the tag sequence is selected from SEQID NO.1 to SEQID NO.18; and the microballons are respectively coated by a specific anti-tag sequence and have codes in different colors. The invention also discloses the SNP detection liquid phase chip for apoE and KIF6 genes and the chromosome 9p21 section. The occlusion rate of the detection method and the sequencing method which are provided in the invention is as high as 100 percent. The prepared SNP detection liquid phase chip for the apoE and KIF6 genes and the chromosome 9p21 section has favorable single-noise ratio.

The invention discloses an AGT1R (Angiotensin Type 1 Receptor) gene SNP (Single Nucleotide Polymorphism) detection specific primer and a liquid phase chip. The liquid phase chip mainly comprises an ASPE (Allele Specific Primer Extension) primer, a microsphere and an amplification primer, wherein the ASPE primer consists of tag sequences at an ends 5' and specific primer sequences specific to SNP sites at ends 3', the specific primer sequences are respectively selected from SEQ ID NO.5, SEQ ID NO.6, and / or SEQ ID NO.7 and SEQ ID NO.8, and the tag sequences are selected from SEQ ID NO.1-SEQ ID NO.4. The prepared AGT1R gene SNP detection liquid phase chip has excellent signal-noise ratio, and cross reaction does not exist between a designed probe and an anti-tag sequence basically. The ASPE primer designed by the invention has excellent specificity, and can accurately differentiate various kinds of mutant sites.

The invention discloses an FGFR1 gene mutation detection specific primer and a liquid chip. The liquid chip mainly comprises ASPE primers consisting of tag sequences at a 5' end and specific primer sequences aiming at target gene mutation sites at a 3' end, microspheres coated with anti-tag sequences and an amplification primer, wherein the specific primer sequences are as follows: SEQ ID NO.5 and SEQ ID NO.6 which aim at a C374T site, and / or SEQ ID NO.7 and SEQ ID NO.8 which aim at a C754A site. The detection liquid chip provided by the invention has the advantages that the matching rate between the detection result and a sequencing method is up to 100%, and the wild-type and mutant parallel detection of multiple mutant sites can be realized.

The invention discloses an RB1 gene mutation detection specific primer and a liquid chip. The liquid chip mainly comprises ASPE primers consisting of tag sequences at a 5' end and specific primer sequences aiming at target gene mutation sites at a 3' end, microspheres coated with anti-tag sequences and an amplification primer, wherein the specific primer sequences are as follows: SEQ ID NO.7 and SEQ ID NO.8 which aim at a C1654T site, SEQ ID NO.9 and SEQ ID NO.10 which aim at a C1666T site, and / or SEQ ID NO.11 and SEQ ID NO.12 which aim at a C1981T site. The detection liquid chip provided by the invention has the advantages that the matching rate between the detection result and a sequencing method is up to 100%, and the wild-type and mutant parallel detection of multiple mutant sites can be realized.

The invention discloses a specificity primer and a liquid phase chip for SNP detection of CYP4F2 gene. The liquid phase chip comprises an ASPE primer formed by the specificity primer aiming at a target gene SNP locus of the tag sequence on the 5' end and 3' end, wherein the specificity primer is SEQ ID NO.13 and SEQ ID NO.14 aiming at the G1297A SNP locus and / or SEQ ID NO.15 and SEQ ID NO.16 aiming at the T34G SNP locus, a microsphere and an amplification primer. The invention also provides a liquid phase chip for SNP detection of CYP4F2 and EPHX1 genes, which comprises the corresponding compositions of the liquid phase chip for the SNP detection of the CYP4F2 and EPHX1 genes, an ASPE primer pair aiming at the G357A, G19512990A, T337C and / or A416G SNP locus of the EPHX1 gene, microsphere and amplification primer. The detection liquid phase chip has an excellent signal-noise ratio, can avoid cross reaction and can realize the parallel detection of a plurality of SNP loca.

The invention discloses BRAP and PSMA6 gene SNP detection specific primers and a liquid phase chip. The liquid phase chip comprises an ASPE primer which is composed of a tag sequence at a 5' terminal and specific primers mutated for a target gene at a 3' terminal, wherein the specific primers respectively comprise a SEQ ID NO.7 and a SEQ ID NO.8 for a BRAP gene A96G SNP locus, a SEQ ID NO.9 and a SEQ ID NO.10 for a BRAP gene A80G SNP locus, and / or a SEQ ID NO.11 and a SEQ ID NO.12 for a PSMA6 gene C157G SNP locus; microspheres coated by an anti-tag sequence; and amplification primers. The coincidence rate between detection results of the BRAP and PSMA6 gene SNP detection liquid phase chip provided by the invention and results of a sequencing method is up to 100%.

The invention discloses a specific primer and a liquid phase chip to test apo E gene and single nucleotide polymorphism (SNP) of the 9p21 segment of chromosome. The liquid phase chip mainly comprises a pair of allele specific primer extension (ASPE) primers. The specific primer has the SEQ ID No.17 and SEQ ID No.18 corresponding to the C609T SNP site, extension primers and / or SEQ ID No. 19 and SEQ ID No.20 corresponding to the T471C SNP site. The tag sequence is from SEQ ID No.1 to SEQ ID No.16. Micro particles which contain specific anti-tag sequence and have different color codes are respectively contained, and the anti-tap sequence is from SEQ ID No.33 to SEQ ID No.48. The invention also discloses a liquid phase chip to test the apo E gene and the SNP of the 9p21 segment of chromosome. The test result of the liquid phase chip fully matches that of a sequencing method. The prepared liquid phase chip to test the apo-E gene and the SNP of the 9p21 segment of chromosome has high signal to noise ratio.

The invention discloses a specific primmer and a liquid phase chip for solute carrier organic anion transporter 1B1 (SLCO1B1) gene signal nucleotide polymorphism (SNP) detection. The liquid phase chip mainly comprises allele specific primer extension (ASPE) primer pairs, microspheres and amplifiers, wherein each ASPE primer consists of a tag sequence at a 5' end and specific primers at a 3' end aiming at an SNP locus; the specific primers comprise SEQ ID NO.11 and SEQ ID NO.12 aiming at T521C SNP locus, SEQ ID NO.13 and SEQ ID NO.14 aiming at T89595C SNP locus, SEQ ID NO.15 and SEQ ID NO.16 aiming at A388G SNP locus, SEQ ID NO.17 and SEQ ID NO.18 aiming at C463A SNP locus, and / or SEQ ID NO.19 and SEQ ID NO.20 aiming at A1929C SNP locus; and the tag sequence is selected from SEQ ID NO.1 to SEQ ID NO.10. The coincidence rate of the liquid phase chip provided by the invention and the detection result of a sequencing method reaches 100 percent. And the prepared liquid phase chip for the SLCO1B1 gene SNP detection has high signal-to-noise ratio.

The invention discloses specific primers and a liquid phase chip for detecting the polymorphism of a CDKAL1 gene. The liquid phase chip mainly comprises specific primers consisting of a tag sequence at a 5' end and specific primer sequences of polymorphic sites of a target gene, microspheres and amplification primers, wherein the specific primer sequences are one or more pairs from SEQ ID No.9 and SEQ ID No.10 for A107C, SEQ ID No.11 and SEQ ID No.12 for G323C, SEQ ID No.13 and SEQ ID No.14 for A210G and SEQ ID No.15 and SEQ ID No.16 for A75G; the tag sequence may be a sequence from SEQ ID No.1 to SEQ ID No.8. The prepared liquid phase chip for detecting the polymorphism of the CDKAL1 gene has a very high signal-to-noise ratio, and the cross reaction of a designed probe and an anti-tae sequence is prevented basically.

The invention discloses a liquid phase chip for detecting mutation of GSTP1 (glutathione S transferase pl) genes, mainly comprising an ASPE primer consisting of a tag sequence at 5' end and a specific primer at 3' end and aiming at a mutant site, micro-balloons and an amplification primer; wherein the specific primer is SEQ ID NO.11 and SEQ ID NO.12 aiming at the A313G mutant site, and SEQ ID NO.13 and SEQ ID NO.14 aiming at the C341T mutant site; the tag sequence is selected from the group of SEQ ID NO.1 to SEQ ID NO.6; and the micro-balloons different color codes are respectively covered with the special anti-tag sequences. The liquid phase chip for detecting mutation of GSTM1(glutathione S transferase mu), GSTT1 (glutathione S transferase theta) and GSTP1 genes in the invention has a great signal-noise ratio; no cross reaction exists between the designed probes and the anti-tag sequences basically; the tag-tag sequence, the selection of the anti-tag tag sequence and the combinationof the tag-tag sequence with the specific ASPE primer can avoid the cross reaction, and realize parallel detection of a plurality of mutant sites.

The invention discloses CYP1A2 gene detection specific primers and a liquid chip. The liquid chip mainly comprises: various ASPE primers composed of 5' terminal tag sequence and 3' terminal specific primer sequences aiming at target gene mutation site, microspheres coated with anti-tag sequence, and amplification primers. The specific primer sequences comprise: SEQ ID NO.15 and SEQ ID NO.16 aiming at G216A site; SEQ IDNO.17 and SEQ ID NO.18 aiming at C96A site; SEQ ID NO.19 and SEQ ID NO.20 aiming at C139T site; SEQ ID NO.21 and SEQ ID NO.22 aiming at G100A T site; SEQ ID NO.23 and SEQ ID NO.24 aiming at A141T site; SEQ ID NO.25 and SEQ ID NO.26 aiming at C113T site; and / or SEQ ID NO.27 and SEQ ID NO.28 aiming at G110A site. The matching rate of the detection result obtained by using the liquid chip provided by the invention and a result obtained by using a sequencing method is 100%. Therefore, wild-type and mutant-type parallel detections of a plurality of mutation sites are realized.

The invention discloses a XRCC2 gene mutation detection liquid chip, and specific primers. The XRCC2 gene mutation detection liquid chip mainly comprises: ASPE primers, wherein each ASPE primer is composed of 5'-terminal tag sequence, and 3'-terminal specific primer sequence targeting target gene mutation sites, and the specific primer sequence comprises SEQ ID No.9 and SEQ ID No.10 targeting G87A site, SEQ ID No.11 and SEQ ID No.12 targeting G158C site, SEQ ID No.13 and SEQ ID No.14 targeting A82G site, and / or SEQ ID No.15 and SEQ ID No.16 targeting A155C site; microballoons coated with anti-tag sequences; and amplification primers. Self-agreement ratio of detection results of the liquid chip with detection results of sequencing is as high as 100%; and parallel detection of wild types and mutant types of a plurality of mutation sites is realized.

The invention discloses an apoA5 genic mutation detection specific primer and a liquid phase chip, wherein the liquid phase chip mainly includes an ASPE primer formed by 5'end tag sequence and 3'end specific primers, and the specific primers are SEQ ID NO.13 and SEQ ID NO.14 for the mutation of A134G, SEQ ID NO.15 and SEQ ID NO.16 for the mutation of T63C, SEQ ID NO.17 and SEQ ID NO.18 for the mutation of C41G, SEQ ID NO.19 and SEQ ID NO.20 for the mutation of G54T, SEQ ID NO.21 and SEQ ID NO.22 for the mutation of A81G, and / or SEQ ID NO.23 and SEQ ID NO.24 for the mutation of G194A; a microsphere enveloped by anti-tag sequence; and an amplimer. The matching degree of the detection result of the apoA5 genic mutation detection liquid phase chip and the sequencing method is 100%, and the parallel detection of multiple polymorphic sites can be achieved.

The invention discloses a liquid chip and specific primer for detecting SNP of a GPIIIa gene. The liquid chip mainly comprises a wild type ASPE primer and a mutant type ASPE primer which are respectively designed aiming at the SNP loci of the GPIIIa gene, microspheres which are respectively coated with specific anti-tag sequences and have different colors of codes and an amplification primer for amplifying the GPIIIa gene target sequence containing the T196C SNP loci, wherein each ASPE primer is formed by the tag sequence at the 5, terminal and the specific primer, aiming at the SNP loci of the target gene, at the 3, terminal; and the anti-tag sequences are selected from SEQ ID NO.17 to SEQ ID NO.24 and correspondingly complement and are paired with the tag sequences selected from each ASPE primer. The invention also provides a liquid chip and specific primer for detecting SNP of COX-1 and GPIIIa genes. Besides the corresponding composition of the GPIIIa gene, the liquid chip also comprises ASPE primer pair aiming at the SNP loci of the COX-1 gene, microspheres coated with specific anti-tag sequences and an amplification primer. The liquid chip has very good signal to noise ratio,can avoid cross reaction and can realize parallel detection of a plurality of SNP loci.

The present invention discloses a detection liquid phase chip and specific primers for FYCO1 gene mutation. The liquid phase chip mainly comprises: ASPE primers comprising a 5' terminal tag sequence and a 3' terminal target gene mutational site-targeted specific primer sequence, wherein the specific primer sequence comprises A92G site-targeted SEQ ID NO.1, A92G site-targeted SEQ ID NO.2, and / or C57T site-targeted SEQ ID NO.3 and C57T site-targeted SEQ ID NO.4; anti-tag sequence coated microspheres; and amplification primers. According to the present invention, coincidence frequency of the detection results of the detection liquid phase chip and the sequencing method is up to 100%, and single and parallel detection on the wild-type with multiple mutational sites and the mutant-type with multiple mutational sites can be achieved.

The invention discloses specific primers and a liquid-phase chip for SNP (Single Nucleotide Polymorphism) detection of MTHFR and FGF5 genes. The liquid-phase chip comprises an ASPE primer, anti-tag sequence coated microspheres, and an amplification primer, wherein the ASPF primer is composed of a 5'-terminal tag sequence and 3'-terminal specific primers for target gene mutation, and the specific primers are respectively SEQ ID NO. 9 and SEQ ID NO. 10 specific for a G121A SNP site, SEQ ID NO. 11 and SEQ ID NO. 12 specific for an A105G SNP site, SEQ ID NO. 13 and SEQ ID NO. 14 specific for a G111A SNP site, and / or SEQ ID NO. 15 and SEQ ID NO. 16 specific for a T196A SNP site of the FGF5 gene. According to the invention, the coincidence rate between the detection result of the liquid-phase chip for SNP detection of the MTHFR and FGF5 gene and the detection result of a sequencing method is up to 100%.

The invention discloses CYP2C8 gene detection specific primers and a liquid chip. The liquid chip mainly comprises: various ASPE primers composed of 5' terminal tag sequence and 3' terminal specific primer sequences aiming at target gene mutation site, microspheres coated with anti-tag sequence, and amplification primers. The specific primer sequences comprise: SEQ ID NO.13 and SEQ ID NO.14 aiming at A127T site; SEQ ID NO.15 and SEQ ID NO.16 aiming at A124G site; SEQ ID NO.17 and SEQ ID NO.18 aiming at G145A site; SEQ ID NO.19 and SEQ ID NO.20 aiming at 204delA site; SEQ ID NO.21 and SEQ ID NO.22 aiming at C115G site; and / or SEQ ID NO.23 and SEQ ID NO.24 aiming at G315C site. The matching rate of the detection result obtained by using the liquid chip provided by the invention and a result obtained by using a sequencing method is 100%. Therefore, wild-type and mutant-type parallel detections of a plurality of mutation sites are realized.

The invention discloses specific primers and a liquid phase chip for SNP (Single Nucleotide Polymorphism) detection of a STK39 (Serine / Threonine Kinase) gene. The liquid phase chip mainly comprises an ASPE (Allele Specific Primer Extension) primer composed of a tag sequence at a 5' side and specific primers aiming at target gene mutation at a 3' side. The specific primers are respectively: SEQ ID NO.7 and SEQ ID NO.8 aiming at the T107C SNP site, SEQ ID NO.9 and SEQ ID NO.10 aiming at the G232A SNP site, and / or SEQ ID NO.11 and SEQ ID NO.12 aiming at the A190C SNP site; a microsphere coated with an anti-tag sequence; and an amplification primer. The coincidence rate of the detection result of the liquid phase chip for SNP detection of the STK39 gene provided by the invention and a sequencing method is as high as 100%. The prepared liquid phase chip has an excellent signal-noise ratio and can realize parallel detection of multiple polymorphism sites.

The invention discloses CYP2A6 gene detection specific primers and a liquid chip. The liquid chip mainly comprises: various ASPE primers composed of 5' terminal tag sequence and 3' terminal specific primer sequences aiming at target gene mutation site, microspheres coated with anti-tag sequence, and amplification primers. The specific primer sequences comprise: SEQ ID NO.17 and SEQ ID NO.18 aiming at G84T site; SEQ IDNO.19 and SEQ ID NO.20 aiming at G133A site; SEQ ID NO.21 and SEQ ID NO.22 aiming at T229A site; SEQ ID NO.23 and SEQ ID NO.24 aiming at T106C site; SEQ ID NO.25 and SEQ ID NO.26 aiming at G148T site; SEQ ID NO.27 and SEQ ID NO.28 aiming at T149G site; SEQ ID NO.29 and SEQ IDNO.30 aiming at T53C site; and / or SEQ ID NO.31 and SEQ ID NO.32 aiming at G108A site. The matching rate of the detection result obtained by using the liquid chip provided by the invention and a result obtained by using a sequencing method is 100%. Therefore, wild-type and mutant-type parallel detections of a plurality of mutation sites are realized.

The invention discloses CYP1A1 gene detection specific primers and a liquid chip. The liquid chip mainly comprises: various ASPE primers composed of 5' terminal tag sequence and 3' terminal specific primer sequences aiming at target gene mutation site, microspheres coated with anti-tag sequence, and amplification primers. The specific primer sequences comprise: SEQ ID NO.17 and SEQ ID NO.18 aiming at T109C site; SEQ ID NO.19 and SEQ ID NO.20 aiming at T97C site; SEQ ID NO.21 and SEQ ID NO.22 aiming at 133-134insT site; SEQ ID NO.23 and SEQ ID NO.24 aiming at T201A site; SEQ ID NO.25 and SEQ ID NO.26 aiming at C240A site; SEQ ID NO.27 and SEQ ID NO.28 aiming at A242G site; SEQ ID NO.29 and SEQ ID NO.30 aiming at C248A site; and / or SEQ ID NO.31 and SEQ ID NO.32 aiming at G135T site. The matching rate of the detection result obtained by using the liquid chip provided by the invention and a result obtained by using a sequencing method is 100%. Therefore, wild-type and mutant-type parallel detections of a plurality of mutation sites are realized.

The invention discloses CYP2B6 gene detection specific primers and a liquid chip. The liquid chip mainly comprises: various ASPE primers composed of 5' terminal tag sequence and 3' terminal specific primer sequences aiming at target gene mutation site, microspheres coated with anti-tag sequence, and amplification primers. The specific primer sequences comprise: SEQ ID NO.13 and SEQ ID NO.14 aiming at C123T site; SEQ ID NO.15 and SEQ ID NO.16 aiming at A101G site; SEQ ID NO.17 and SEQ ID NO.18 aiming at G118T site; SEQ ID NO.19 and SEQ ID NO.20 aiming at A77G site; SEQ ID NO.21 and SEQ IDNO.22 aiming at T139C site; and / or SEQ ID NO.23 and SEQ ID NO.24 aiming at C288T site. The matching rate of the detection result obtained by using the liquid chip provided by the invention and a result obtained by using a sequencing method is 100%. Therefore, wild-type and mutant-type parallel detections of a plurality of mutation sites are realized.