TaqMan probe real-time fluorescence PCR method for detecting rs6313 locus of HTR2A gene and primer probe combination

A primer probe, real-time fluorescence technology, applied in biochemical equipment and methods, DNA/RNA fragments, recombinant DNA technology, etc., can solve the problems of patients not responding to treatment, achieve easy analysis of experimental results, save time and consumables, Simple operation effect

Pending Publication Date: 2020-01-03
SHANXI LIFEGEN
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Despite the wide variety of medications, approximately 30-50% of patients do not respond to treatment

Method used

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  • TaqMan probe real-time fluorescence PCR method for detecting rs6313 locus of HTR2A gene and primer probe combination
  • TaqMan probe real-time fluorescence PCR method for detecting rs6313 locus of HTR2A gene and primer probe combination
  • TaqMan probe real-time fluorescence PCR method for detecting rs6313 locus of HTR2A gene and primer probe combination

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Experimental program
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specific Embodiment

[0042] Specific embodiment: TaqMan probe real-time fluorescent PCR method detects the allele of rs6313 site.

[0043] 1. Extraction and dilution of DNA samples

[0044] After collecting venous blood in vacuum blood collection tubes anticoagulated with ethylenediaminetetraacetic acid (EDTA) according to conventional methods, DNA was extracted using the QIAamp DNA Mini Blood Kit (Qiagen, Germany) kit; the extracted DNA was concentrated using NanoDrop 2000 Determination (A260 / 280=1.95~2.15). Using the above method, respectively measure the concentration of 104 cases of Bouyei DNA samples, and then use PCR grade H 2 O Dilute the sample to 10 ng / μl.

[0045] 2. Design primers and probes

[0046] Specific upstream primers: C allele: 5'-GCCTTACAGTAATGACTTTAACTTC-3'; T allele: 5'-GCCTTACAGTAATGACTTTAACTTT-3';

[0047] Universal downstream primer: 5'-GATGAAGTAAGGAGAGACACGAC-3';

[0048] Probe: 5'-FAM-TAACACTTCTGATGCATTTAACTGGAC-BHQ2-3';

[0049] Among them, FAM is 6-carboxyfluore...

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Abstract

The invention belongs to the field of gene diagnosis, and discloses a TaqMan probe real-time fluorescent PCR method for detecting the rs6313 locus (HTR2A, 102C>T) of a HTR2A gene, and a primer probe combination. A upstream primer FpC-T is 5'-GCTCTACAGTAATGACTTTAACTTC-3', a upstream primer FpT-T is 5'-GCTCTACAGTAATGACTTTAACTTT-3', a downstream primer Rp is 5'-GATGAAGTAAGGAGAGACACGAC-3', and a probeProbe is 5'-FAM-TAACACTTCTGATGCATTTAACTGGAC-BHQ2-3'. After PCR reaction is completed, the presence or absence of an amplification curve can be used for analyzing the result. The TaqMan probe real-time fluorescent PCR method has the advantages of simple operation, high resolution, no pollution, high throughput and the like.

Description

technical field [0001] The invention belongs to the field of pharmacogenomics and gene detection, and specifically relates to the design of a specific primer and probe for the allele of the rs6313 site of the HTR2A gene, and a detection method. Background technique [0002] The latest statistics released by the World Health Organization (WHO) show that the total number of patients with depression worldwide is about 322 million, and the proportion of the global population of patients with depression is estimated to be 4.4%. The prevalence of depression in China is 4.2%. It is conservatively estimated that there are more than 58 million people suffering from depression in China. Depression has become the fourth largest disease in the world at present, and antidepressant drugs such as fluoxetine, paroxetine, and fluvoxamine are the main treatments for depression. Despite the wide variety of drugs, approximately 30-50% of patients do not respond to treatment. A randomized cont...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6858C12N15/11
CPCC12Q1/6858C12Q2535/137C12Q2561/101C12Q2531/113C12Q2545/101
Inventor 王会娟康星韩敏
Owner SHANXI LIFEGEN
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