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Application of exosomal microRNA molecular markers and kits for diagnosing esophageal cancer

A technology of molecular markers and exosomes, which is applied in the field of medical molecular biology, can solve the problems of insufficient detection specificity of microRNA molecular markers, and achieve the effects of high reverse transcription efficiency, strong primer specificity, and high sensitivity

Active Publication Date: 2022-03-01
JIANGSU MICRODIAG BIOMEDICINE TECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0009] The second purpose of the present invention is to provide a kit for diagnosing esophageal cancer, so as to alleviate the technical problem in the prior art that the detection specificity of microRNA molecular markers related to esophageal cancer exosomes is not strong enough

Method used

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  • Application of exosomal microRNA molecular markers and kits for diagnosing esophageal cancer
  • Application of exosomal microRNA molecular markers and kits for diagnosing esophageal cancer
  • Application of exosomal microRNA molecular markers and kits for diagnosing esophageal cancer

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0069] Example 1 Two-step method detection system kit and use based on PCR platform miRNA

[0070] 1. RNA reverse transcription reaction system:

[0071]Reagents: The reagents used to prepare the reverse transcription reaction system include reverse transcription primers (RT-Primer, synthesized by Shanghai Yingweijieji), miRNA standard powder (synthesized by Shanghai Yingweijieji), T4 DNA ligase (T4 DNA Ligase, Supplier: NEB, product number: M0202S, including 10×T4 DNA Ligase Buffer), RNase inhibitor (RNase inhibitor, supplier: Fermentas, product number: K1622), transcriptase Transcriptase (supplier: Shanghai Yingwei Jieji Biotechnology Co., Ltd., product number: K1622, containing RNase inhibitor, dNTPs, nuclease-free water), T4 polynucleotide kinase (T4 Polynucleotide Kinase, supplier: NEB, product number: M0201S) and nuclease-free pure water (nuclease-free water, supplier: Shanghai Yingwei Jieji Biological Co., Ltd., product number: K1622). The reagents used to prepare the...

Embodiment 2

[0096] Example 2 Effect Evaluation of Esophageal Cancer Auxiliary Diagnosis Detection Kit Based on the Two-step Detection System of miRNA

[0097] 1. Sample collection

[0098] Tissue, serum, plasma, and urine samples were collected from a series of people including esophageal cancer (including different stages, different subtypes, different genders, and different age groups), benign lesions of esophageal diseases, and healthy people diagnosed by hospital examination.

[0099] 2. Tissue miRNA extraction and purification

[0100] Using QIAGEN’s commercial product miRNeasy Serum / Plasma Kit (Product No. 217184), extract and purify miRNA in tissue and serum, and use Nano-Drop 2000 to measure RNA nucleic acid quality, record RNA concentration and purity, and normalize tissue miRNA One treatment.

[0101] 3. Extraction and purification of serum exosomal miRNA and plasma exosomal miRNA

[0102] Serum and plasma exosomes were extracted using the commercial product ExoQuickTM kit (p...

Embodiment 3

[0134] Example 3 Exosome miRNA expression level and its difference in body fluid before and after esophageal cancer

[0135] 1. Collect esophageal cancer diagnosed by the hospital (including different stages, different subtypes, different genders and different age groups), and collect preoperative body fluid (serum, plasma, urine) samples from 10 patients with esophageal cancer who have not received any treatment And corresponding postoperative body fluid (serum, plasma, urine) samples. The expression levels of miR-21, miRNA-223, miRNA-25, miRNA-205, miR-203a, miR-375, mi-R-146b-5p, miR-194, and miR-145 were detected.

[0136] 2. The miRNA markers miR-21, miR-223, miR-25, miR-205, miR-375, and miR-203a were detected in clinical samples, and the preoperative body fluids (serum, Plasma, urine) samples and corresponding postoperative body fluid (serum, plasma, urine) samples for Exo-miRNA detection, detection of miRNA markers miR-21, miR-223, miR-25, miR-205, miR- 375, the expr...

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Abstract

The present invention provides an application of exosomal microRNA molecular markers and a kit for diagnosing esophageal cancer, which relate to the technical field of medical molecular biology. The microRNA molecular markers provided by the present invention are found in patients with esophageal cancer The expression of exosomes in body fluids changed significantly. Therefore, the microRNA molecular markers in body fluid exosomes provided in the present invention can be used as markers for risk estimation and detection of esophageal cancer, which is very helpful for the prevention and treatment of esophageal cancer in my country. In addition, the present invention also provides a kit for diagnosing esophageal cancer, including primers and probes for detecting microRNA molecular markers. The reverse transcription efficiency is high, the primer specificity is strong, the sensitivity is high, the detection is comprehensive, fast and accurate, and the application is wide, and the detection results can be used for clinical guidance of medication.

Description

technical field [0001] The invention relates to the technical field of medical molecular biology, in particular to the application of an exosome microRNA molecular marker and a kit for diagnosing esophageal cancer. Background technique [0002] Esophageal cancer is one of the common digestive tract tumors, ranking sixth among malignant tumors in the world. In my country, the incidence and mortality of malignant tumors rank fifth and fourth respectively. But from a global perspective, the incidence of esophageal cancer in my country ranks first in the world, and about half of the patients who die from esophageal cancer every year are Chinese (about 300,000 people die from esophageal cancer every year in the world, and the average death rate in my country is about 150,000 people). Most of them have reached the middle and late stages when they seek medical treatment, and the treatment costs are relatively high. The key to its prevention and treatment lies in early detection, ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/6886C12Q1/6851
CPCC12Q1/6851C12Q1/6886C12Q2600/178C12Q2600/158C12Q2600/118C12Q2531/113C12Q2521/107C12Q2525/301C12Q2545/101
Inventor 王弢渠香云董肇楠马雪情贾云莉
Owner JIANGSU MICRODIAG BIOMEDICINE TECH CO LTD
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