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Specific primer pair, reagent and kit for detecting a gene C4BPA mRNA in human peripheral blood leukocytes, and applications of specific primer pair, reagent and kit

A peripheral blood and leukocyte technology, applied in DNA/RNA fragments, recombinant DNA technology, biochemical equipment and methods, etc., can solve problems such as limited clinical diagnostic value or predictive significance

Inactive Publication Date: 2021-07-23
NANJING FIRST HOSPITAL
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Foreign scholars Fontana P (2013) reported that the sensitivity of CYP2C19 genotyping to predict clopidogrel resistance is only 37.6%, which is close to our data of 30.4%, showing that its clinical diagnostic value or predictive significance is very limited

Method used

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  • Specific primer pair, reagent and kit for detecting a gene C4BPA mRNA in human peripheral blood leukocytes, and applications of specific primer pair, reagent and kit
  • Specific primer pair, reagent and kit for detecting a gene C4BPA mRNA in human peripheral blood leukocytes, and applications of specific primer pair, reagent and kit
  • Specific primer pair, reagent and kit for detecting a gene C4BPA mRNA in human peripheral blood leukocytes, and applications of specific primer pair, reagent and kit

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0018] Example 1. The application of the mRNA expression level of C4BPA in human peripheral blood leukocytes in the preparation of a kit for detecting clopidogrel resistance.

[0019] 1. A kit for detecting clopidogrel resistance, including the following components:

[0020] TRIzol reagent;

[0021] Chloroform;

[0022] isopropanol;

[0023] DEPC water

[0024] gDNA eraser

[0025] 5×gDNA eraser buffer

[0026] RT enzyme mix I

[0027] 5× buffer2 (for Real-Time)

[0028] RT primer mix

[0029] RNase-free dH 2 o

[0030] Premix Ex Taq TM II(2×)

[0031] ROX Reference Dye II (50×)

[0032] Specific primer sequences for detection of C4BPA mRNA are SEQ ID NO.1 and SEQ ID NO.2.

[0033] The primer pair (SEQ ID NO.3 and SEQ ID NO.4) of internal reference GAPDH required for mRNA quantitative analysis.

[0034] 2. How to use the kit

[0035] The total RNA of peripheral blood leukocytes was extracted, and the mRNA expression level of C4BPA was detected by qRT-PCR t...

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Abstract

The invention discloses a specific primer pair, reagent and kit for detecting a gene C4BPA mRNA in human peripheral blood leukocytes, and applications of the specific primer pair, reagent and kit. A nucleotide sequence of an upstream primer of the primer pair for detection is shown as SEQ ID No. 1, and the nucleotide sequence of a downstream primer is shown as SEQ ID No. 2. Clinically, through the detection of C4BPA mRNA in the human peripheral blood leukocytes, clopidogrel resistance can be predicted, so that a novel detection method can be provided for judging the clinical effects and prognosis of clopidogrel and guiding clinical rational use of drugs.

Description

[0001] This application is a divisional application with an application date of April 1, 2019, an application number of 201910255608.1, and an invention title of "Detection of Human Peripheral Blood Cell Complement Inhibitor C4BPA mRNA Detection and Prediction of Clopidogrel Resistance and Preparation of a Kit". The applicant is Nanjing First Hospital. technical field [0002] The invention belongs to the field of biological detection, and relates to a pair of specific primers, a reagent and a kit for detecting gene C4BPAmRNA in human peripheral blood leukocytes and applications thereof. Background technique [0003] As a classic antiplatelet drug, clopidogrel is commonly used in stroke, acute coronary syndrome (ACS) and percutaneous coronary intervention (PCI) implantation of drug-eluting stents (to prevent stent thrombosis), The "dual antiplatelet therapy" combined with aspirin is the "gold standard" antiplatelet therapy after ACS or PCI (stent implantation), and has been ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6883C12N15/11
CPCC12Q1/6883C12Q2600/158C12Q2600/106
Inventor 谢红光吉金子黄贝贝邰婷米琼宇谢文剑
Owner NANJING FIRST HOSPITAL
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