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Human histamine receptor HRH1 mRNA detection primer probe set, kit and application

A histamine receptor, primer probe technology, applied in DNA/RNA fragments, recombinant DNA technology, biochemical equipment and methods, etc., can solve the problems of lack of research and records, unable to quantify treatment effects, etc., to achieve rapid dynamic monitoring and efficacy assessment, reducing the risk of aerosol contamination, increasing the effect of accuracy

Pending Publication Date: 2021-11-05
ZHEDA DIXUN BIO GENE ENG
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  • Description
  • Claims
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AI Technical Summary

Problems solved by technology

[0004] Most patients with allergic diseases are treated with symptomatic treatment based on experience, and antihistamines are used to reduce allergic symptoms, but antihistamine treatment is not effective for all patients, and the therapeutic effect cannot be quantified, but there is no relevant research research and documentation

Method used

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  • Human histamine receptor HRH1 mRNA detection primer probe set, kit and application
  • Human histamine receptor HRH1 mRNA detection primer probe set, kit and application
  • Human histamine receptor HRH1 mRNA detection primer probe set, kit and application

Examples

Experimental program
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Effect test

Embodiment 1

[0057] 1. Entrust Shanghai Sunny Biotechnology Co., Ltd. to synthesize the primers and probes shown in Table 1.

[0058] 2. Standard product preparation

[0059] In vitro transcription. Using pGM-T ligation kit [Tiangen Biochemical Technology (Beijing) Co., Ltd., article number: VT202-01], using pGM-T as the vector to construct HRH1 plasmid DNA (commissioned by Nanjing GenScript Co., Ltd. to construct and synthesize, Figure 8 ), HRH1 plasmid DNA was transcribed into mRNA in vitro with HiScribe T7 High Yield RNA Synthesis Kit (produced by NEW ENGLAND BioLabs, Cat. No.: E2040S).

[0060] According to the copy number calculation formula: copy number=[6.02×10 23 × RNA concentration (ng / μL) × 10 -9 ] / [RNA length (bp)×340] to calculate the initial RNA copy number. Dilute to 1.0 x 10 with nuclease-free water 10 copeies / μL, which is HRH1 standard.

[0061] 3. Whole blood RNA extraction and dilution: EDTA anticoagulated whole blood samples were extracted with a whole blood total...

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Abstract

The invention provides a human histamine receptor HRH1 mRNA detection primer probe set, a kit and application, and relates to the technical field of biological detection. The primer probe group disclosed by the invention comprises HRH1-F, HRH1-R and a probe H1-Probe, the nucleotide sequence of the HRH1-F is as shown in SEQ ID NO.1, the nucleotide sequence of the HRH1-R is as shown in SEQ ID NO.2, and the nucleotide sequence of the probe H1-Probe is as shown in SEQ ID NO.3. The invention provides a kit containing the primer probe group and a detection method, the expression level of HRH1 mRNA can be detected by using an RNA one-step method, and a detection means with high accuracy, wide detection range and high sensitivity is provided for detection of HRH1 protein.

Description

technical field [0001] The invention belongs to the technical field of biological detection, and in particular relates to a human histamine receptor HRH1 mRNA detection primer probe set, kit and application. Background technique [0002] Histamine is an active amine compound, a biogenic amine widely present in animals and plants, formed by decarboxylation of histidine, and usually stored in mast cells of tissues. It is an important chemical transduction substance in the body, which can affect the reactions of many cells, including allergies, inflammatory reactions, gastric acid secretion, etc. When the body is stimulated by a certain stimulus to trigger an antigen-antibody reaction, it will cause the cell membrane of mast cells to permeate Sex changes, releases histamine, and interacts with histamine receptors to produce pathophysiological effects. Histamine synthesis occurs mainly in mast cells, basophils, lungs, skin, and gastrointestinal mucosa, consistent with histamine...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6883C12N15/11
CPCC12Q1/6883C12Q2600/106C12Q2600/158
Inventor 吴善东刘奕吴周杰蒋学翰王吉萍王美杰杨旭凯
Owner ZHEDA DIXUN BIO GENE ENG
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