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Primers, probe, fluorescent PCR kit and method for detecting human HLA-B*1301 gene

A technology for HLA-B and detection primers, which is applied in the field of biomedical clinical molecular detection, can solve the problems of high price, long cycle, cumbersome data analysis, etc., and achieve the effect of ensuring authenticity and accuracy, low cost and wide promotion value

Inactive Publication Date: 2015-03-25
SUZHOU KUANGYUAN MOLECULAR BIOTECH
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] The present invention aims at the disadvantages of the prior art, such as high price, long period, low efficiency, cumbersome data analysis, etc. when detecting HLA-B*1301 alleles, and provides a HLA-B*1301 based on Taqman real-time fluorescent quantitative PCR technology platform Primers, fluorescent probes, kit and detection method for allele detection, used for detection of human HLA-B*1301 allele

Method used

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  • Primers, probe, fluorescent PCR kit and method for detecting human HLA-B*1301 gene
  • Primers, probe, fluorescent PCR kit and method for detecting human HLA-B*1301 gene
  • Primers, probe, fluorescent PCR kit and method for detecting human HLA-B*1301 gene

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0079] Example 1. Design of primers and probes:

[0080] According to the reference sequence of the HLA-B gene (NG_023187) released by the nucleotide sequence database GeneBank of the National Center for Biotechnology Information NCBI (NG_023187) and the information of HLA-B*1301 (HLA00152) published by the British IMG / HLA database, Primer Express of ABI Company was used 3.0 software respectively designed primers and probes for detecting HLA-B*1301 and other genes. Primers and probes, as follows:

[0081] Forward primer:

[0082] HLA-B*1301 F: 5’-GGGCCAGGGTCTCACATCA-3’

[0083] Reverse primer:

[0084] HLA-B*1301 R: 5’-CACGGGCCGCCTCCCACTTGA-3’

[0085] Fluorescent probes:

[0086] TM-HLA-B*1301: 5’Fam- CCAGGTAGGCTCTCAGCTGCTCCG-3’BHQ1

[0087] In order to monitor the effectiveness of the reaction system, internal control primers and probes are added to the detection system. The present invention selects a sequence of the human conserved gene GAPDH (its reference sequence ...

Embodiment 2

[0093] Example 2. Preparation of primers

[0094] The designed primers and probe sequences are handed over to the synthesis company for synthesis, generally using automatic chemical synthesis with instruments, and a synthesis inspection report is required.

Embodiment 3

[0095] Example 3: Preparation of a kit for detecting human HLA-B*1301 alleles by fluorescent PCR

[0096] Prepare a reaction solution for detecting human HLA-B*1301 alleles, in which the PCR reaction solution contains not only buffers, magnesium ions, dNTPs and other substances necessary for PCR reactions, but also detection primers and probes and internal control primers and probes. The concentration of the PCR reaction solution components and the sequence information of the included primers and probes for the above-mentioned specific detection are as follows:

[0097] Table 1. Components of PCR reaction solution

[0098]

[0099] HLA-B*1301 allele

[0100] HLA-B*1301 F: 5’-GGGCCAGGGTCTCACATCA-3’

[0101] HLA-B*1301 R: 5’-CACGGGCCGCCTCCCACTTGA-3’

[0102] TM-HLA-B*1301: 5’Fam- CCAGGTAGGCTCTCAGCTGCTCCG-3’BHQ1

[0103] GAPDH F: 5'-GCTGCTTTTTAACTCTGGTAAAGTG-3';

[0104] GAPDH R: 5'-TAGCACTCACCATGTAGTTGAG-3';

[0105] TM-GAPDH: 5'Rox-TGATGCATCTATGAACGCTTC-3'BHQ2.

[010...

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Abstract

The invention provides primers, a probe, a fluorescent PCR kit and a detection method for detecting a human HLA-B*1301 allele. According to the theory of 'allele specific PCR', a human leucocyte antigen HLA-B*1301 allele can be detected on a real-time fluorescent quantitative PCR technical platform.

Description

technical field [0001] The invention belongs to the field of biomedical clinical molecular detection, in particular to a primer, a probe, a fluorescent PCR kit and a detection method for detection of an HLA allele type HLA-B*1301 allele. Background technique [0002] Human leukocyte antigen (HLA) is a group of genes located on chromosome 6. It is the gene complex with the highest allelic polymorphism among known genes so far. More than 2,700 alleles have been found in the world, although most of them Alleles are very rare. HLA gene is the most complex genetic polymorphism system in humans, and its polymorphism distribution has obvious ethnic characteristics. Studies have shown that there is a strong correlation between the HLA-B*1301 allele and the "dapsone syndrome" caused by the drug dapsone. [0003] Dapsone is a sulfone antibacterial agent, which has a strong antibacterial effect on Bacillus leprae. Its mechanism of action is similar to that of sulfa drugs, acting on ...

Claims

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Application Information

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IPC IPC(8): C12Q1/68C12N15/11
CPCC12Q1/6883C12Q1/6844C12Q2600/156C12Q2600/166
Inventor 王进吴梦华赵小龙吴子侠石亦静
Owner SUZHOU KUANGYUAN MOLECULAR BIOTECH
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