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Application of miR-493-5p detection reagent in preparation of esophageal cancer detection kit and esophageal cancer detection kit

A detection kit and detection reagent technology, applied in the fields of biotechnology and medicine, can solve the problems of inability to diagnose esophageal cancer, low specificity, and large patient trauma, and achieve high clinical use value and promotion value, good specificity, highly specific effect

Active Publication Date: 2021-02-12
THE SECOND HOSPITAL OF SHANDONG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Gastroscopy is currently the main method for diagnosing esophageal cancer, but it is traumatic to the patient and requires high technical skills for the operator, so it is not suitable for large-scale population screening
Upper gastrointestinal barium meal, CT and other examinations cannot be used for the diagnosis of esophageal cancer because of their low specificity.

Method used

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  • Application of miR-493-5p detection reagent in preparation of esophageal cancer detection kit and esophageal cancer detection kit
  • Application of miR-493-5p detection reagent in preparation of esophageal cancer detection kit and esophageal cancer detection kit
  • Application of miR-493-5p detection reagent in preparation of esophageal cancer detection kit and esophageal cancer detection kit

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0033] Example 1, the expression level of miR-493-5p in plasma exosomes of patients with early esophageal cancer

[0034] 1. Research objects: 30 patients with early esophageal cancer (according to postoperative pathological TNM stage: T1N0M0) admitted to the Second Hospital of Shandong University from August 2019 to June 2020 were included as the experimental group, and 30 healthy volunteers were included as the experimental group. control group. None of the included patients with esophageal cancer received neoadjuvant chemoradiotherapy before operation.

[0035] 2. Plasma collection: Collect about 3mL of peripheral blood from patients with early esophageal cancer before operation, place them in a vacuum tube containing EDTA anticoagulant and mix them upside down, centrifuge at 1900g, 4°C for 10min, and collect the supernatant; Centrifuge again at ℃ for 15 minutes and carefully pipette the supernatant into a cryopreservation tube, freeze it in liquid nitrogen and store it in...

Embodiment 2

[0060] Example 2. Differences in the expression level of miR-493-5p between patients with and without lymph node metastasis of esophageal cancer

[0061] The qPCR method was used to quantify the level of miR-493-5p in the preoperative plasma exosomes of 30 groups of esophageal cancer patients with lymph node metastasis and 30 groups of esophageal cancer patients without lymph node metastasis detected by postoperative pathology, so as to determine the level of plasma exosomes. Whether miR-493-5p can be used as a biomarker to judge whether there is lymph node metastasis in patients with esophageal cancer.

[0062] Collect the preoperative plasma of esophageal cancer patients with and without lymph node metastasis, separate plasma exosomes, extract the total RNA of plasma exosomes, measure the concentration of the extracted total RNA, and use the tailing method to reverse transcribe to obtain cDNA, and the obtained cDNA Real-time fluorescent quantitative PCR was carried out, all ...

Embodiment 3

[0066] Example 3. Differences in miR-493-5p expression levels in esophageal cancer plasma exosomes before and after surgery

[0067] The plasma of patients with esophageal cancer before operation and 7 days after operation was collected, plasma exosomes were separated, total RNA of plasma exosomes was extracted, the concentration of the extracted total RNA was measured, cDNA was obtained by reverse transcription by tailing method, and the obtained cDNA Real-time fluorescent quantitative PCR was carried out, all of which were carried out according to the operation method in Example 1.

[0068] Real-time fluorescence quantitative results such as Figure 6 As shown, by comparing the expression of miR-493-5p in plasma exosomes of patients with esophageal cancer before and after surgery, it was found that the expression level of miR-493-5p in plasma exosomes of patients with esophageal cancer after surgery was significantly higher than that before surgery. High (p<0.001). The kit...

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Abstract

The invention relates to an application of a miR-493-5p detection reagent in preparation of an esophageal cancer detection kit and the esophageal cancer detection kit. The invention reveals for the first time that miR-493-5p in plasma exosome is related to esophageal cancer and lymph node metastasis of esophageal cancer, is significantly reduced in patients with esophageal cancer and lymph node metastasis, and has statistical significance. On the basis, the miR-493-5p detection reagent is applied to preparation of the esophageal cancer detection kit, and the prepared kit can be used for conveniently and quickly carrying out early diagnosis of esophageal cancer, early warning of lymph node metastasis and judgment of operative treatment conditions of esophageal cancer patients with high specificity, so guidance is provided for treatment of the esophageal cancer patients.

Description

technical field [0001] The invention relates to the application of a miR-493-5p detection reagent in the preparation of an esophageal cancer detection kit and the esophageal cancer detection kit, belonging to the fields of biotechnology and medical technology. Background technique [0002] Esophageal cancer is a common malignant tumor of the digestive tract, and its incidence rate ranks eighth among malignant tumors worldwide, and its mortality rate ranks sixth. my country is an area with a high incidence of esophageal cancer, and about 90% of them are esophageal squamous cell carcinoma. In recent years, although the diagnosis and treatment technology of esophageal cancer has been improved, its prognosis is still poor, and the 5-year overall survival rate is only about 20%. Due to the concealment and non-specificity of the early symptoms of esophageal cancer, most patients are already in the middle and late stage after clinical diagnosis and treatment. Gastroscopy is curre...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6886C12Q1/6851C12N15/11
CPCC12Q1/6886C12Q1/6851C12Q2600/178C12Q2600/118C12Q2600/158C12Q2531/113C12Q2545/101C12Q2563/107
Inventor 赵小刚肖兆华李佩蔚田忠献姜宁周洁罗钧文
Owner THE SECOND HOSPITAL OF SHANDONG UNIV
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