Primer group and kit for detecting mRNA expression of cationic protein of human eosinophilic granulocyte

A technology of eosinophils and cationic proteins, which is applied in the direction of recombinant DNA technology, DNA/RNA fragments, biochemical equipment and methods, and can solve the problems of small detection range, accuracy problems, and low sensitivity

Pending Publication Date: 2021-09-10
ZHEDA DIXUN BIO GENE ENG
View PDF8 Cites 1 Cited by
  • 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

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Primer group and kit for detecting mRNA expression of cationic protein of human eosinophilic granulocyte
  • Primer group and kit for detecting mRNA expression of cationic protein of human eosinophilic granulocyte
  • Primer group and kit for detecting mRNA expression of cationic protein of human eosinophilic granulocyte

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0054] 1. Entrust Shanghai Sunny Biotechnology Co., Ltd. to synthesize the primers and probes shown in Table 1.

[0055] 2. Standard product preparation

[0056] In vitro transcription. Using pGM-T ligation kit [Tiangen Biochemical Technology (Beijing) Co., Ltd., article number: VT202-01], using pGM-T as the carrier to construct ECP plasmid DNA (commissioned by Nanjing GenScript Co., Ltd. to construct and synthesize, Figure 8 ), the ECP plasmid DNA was transcribed into mRNA in vitro with HiScribe T7 High Yield RNA Synthesis Kit (manufactured by NEW ENGLAND BioLabs, Cat. No.: E2040S).

[0057] 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 9 copeies / μl, which is the ECP standard.

[0058] 3. Whole blood RNA extraction and dilution

[0059] EDTA anticoagulated whole blood samples were extracted with a...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

PUM

No PUM Login to view more

Abstract

The invention provides a primer group and a kit for detecting mRNA (messenger Ribonucleic Acid) expression of cationic protein of human eosinophilic granulocyte, and relates to the technical field of biological detection. The primer group disclosed by the invention comprises an ECP-F, an ECP-R, a GAPDH-F and a GAPDH-R; the nucleotide sequence of the ECP-F is as shown in SEQ ID NO. 1; the nucleotide sequence of the ECP-R is as shown in SEQ ID NO. 2; the nucleotide sequence of the GAPDH-F is as shown in SEQ ID NO. 3; and the nucleotide sequence of the GAPDH-R is as shown in SEQ ID NO. 4. The invention further provides a kit comprising the above primer group, the expression level of human ECP mRNA can be quantitatively detected by a one-step method, 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 belongs to the technical field of biological detection, and in particular relates to a set of primers and a kit for detecting the expression of human eosinophil cationic protein mRNA. Background technique [0002] Eosinophil cationic protein (eosinophil cationic protein, ECP) ​​is a strongly basic granule protein released by eosinophil (EOS) activation, and exists in the matrix part of EOS granules, accounting for 30% of the granules. ECP is a specific marker of EOS activation, which can reflect the degree of EOS activation, and it also has a strong cytotoxic effect. ECP can be found in most body fluids of the human body, such as serum, nasal secretions, bronchial flushing fluid, saliva, tears, etc. [0003] For inflammatory diseases associated with eosinophil infiltration, the amount of EOS degranulation protein is more important than the total amount of EOS infiltration. EOS granule proteins mainly include: ECP, EOS major basic protei...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

Application Information

Patent Timeline
no application Login to view more
IPC IPC(8): C12Q1/6883C12Q1/6851C12N15/11
CPCC12Q1/6883C12Q1/6851C12Q2600/106C12Q2600/158C12Q2600/166C12Q2531/113C12Q2521/107C12Q2563/107C12Q2545/113C12Q2527/127
Inventor 程雷沈华浩吴善东刘奕吴周杰蒋学翰王教峰王溢飞
Owner ZHEDA DIXUN BIO GENE ENG
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Try Eureka
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap