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Primer and probe set and kit for human beta-tryptase mRNA RT-PCR detection

A technology of tryptase and primer probe, which is applied in the field of biological detection, can solve the problems of accuracy, small detection range, and low sensitivity, and achieve the effect of increasing accuracy, high sensitivity, rapid dynamic monitoring and

Pending Publication Date: 2021-10-29
ZHEDA DIXUN BIO GENE ENG
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The ELISA method has the problems of small detection range and low sensitivity in the detection process, and its accuracy also has problems

Method used

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  • Primer and probe set and kit for human beta-tryptase mRNA RT-PCR detection
  • Primer and probe set and kit for human beta-tryptase mRNA RT-PCR detection
  • Primer and probe set and kit for human beta-tryptase mRNA RT-PCR detection

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0036] 1. The reagents and equipment involved are as follows:

[0037] 1.1 Reagents

[0038] 1.1.1 Whole blood total RNA kit (Hangzhou Xinjing Biological Reagent Development Co., Ltd., catalog number: 5201050)

[0039] 1.1.2 HiScribe T7 HighYield RNA Synthesis Kit (New England Biolabs, Cat. No.: E2050S)

[0040] 1.2 Main instruments

[0041] 1.2.1 Applied Biosystems TM 7300 fluorescent quantitative PCR instrument: ThermoFisher, USA

[0042] 1.2.2 -80℃ low temperature refrigerator: ThermoFisher, USA

[0043] 1.2.3 High-speed and low-temperature desktop centrifuge: Eppendorf, Germany

[0044] 1.2.4 Qubit 3 Fluorometer: ThermoFisher, USA

[0045] 2. Method

[0046] 2.1 Primer and probe design

[0047] According to the sequences of TPSB and GAPDH, using Primer 6.0 software, design fluorescent quantitative primers and probes, after a series of effect verification, the primer pairs TPSB-F, TPSB-R, GAPDH-F, GAPDH-R and probes of TPSB and GAPDH were obtained T-Probe, G-Probe ...

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Abstract

The invention relates to a primer and probe set and kit for human beta-tryptase mRNART-PCR detection, and belongs to the technical field of biological detection. The primer and probe set comprises a primer TPSB-F, a primer TPSB-R and a probe T-Probe, wherein the nucleotide sequence of the primer TPSB-F is shown as SEQ ID NO.1, the nucleotide sequence of the primer TPSB-R is shown as SEQ ID NO.2, and the nucleotide sequence of the probe T-Probe is shown as SEQ ID NO.3. The TaqMan real-time fluorescent quantitative one-step RT-PCR detection primer and probe set is established for human TPSB, and a detection means with high accuracy, wide detection range and high sensitivity is provided for detection of the protein.

Description

technical field [0001] The invention relates to the technical field of biological detection, in particular to a primer probe set and kit for human β-tryptase mRNA RT-PCR detection. Background technique [0002] Mast cells are activated during anaphylaxis, releasing inflammatory mediators including tryptase, a process that contributes to the signs and symptoms of anaphylaxis. Tryptase has the functions of promoting the repair of airway, regulating the tension and reactivity of airway smooth muscle cells, and stimulating the activation of mast cells. At present, tryptase is divided into three categories at the cDNA and protein levels: α, β, γ, among which β has the highest content. After an allergic reaction, a transient increase in the level of β-tryptase (TPSB) in the blood circulation helps to identify and assess the degree of the allergic reaction. 15 minutes to 3 hours of collection. Transiently elevated levels of beta-tryptase in the blood suggest an allergic reaction...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6851C12N15/11
CPCC12Q1/6851C12Q2521/107C12Q2531/113C12Q2563/107C12Q2561/101C12Q2527/127C12Q2545/101C12Q1/6883C12Q2600/158
Inventor 吴善东程雷刘奕吴周杰蒋学翰雷薇王吉萍
Owner ZHEDA DIXUN BIO GENE ENG