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Gastric cancer biomarker based on peripheral blood and detection method and application thereof

A biomarker and peripheral blood technology, applied in the field of gastric cancer biomarkers and their detection, can solve the problems of poor specificity, low sensitivity, and large invasiveness of gastroscopic biopsy, achieve excellent performance, improve sensitivity and specificity, and improve sensitivity and specific effects

Active Publication Date: 2019-11-22
NANJING UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, gastroscopic biopsy is invasive, and commonly used clinical serum tumor-related markers such as carcinoembryonic antigen (CEA), carbohydrate antigen 199 (CA199), carbohydrate antigen 224 (CA224), and carbohydrate antigen 724 (CA724) have low sensitivity , poor specificity and other shortcomings, not suitable for large-scale screening work

Method used

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  • Gastric cancer biomarker based on peripheral blood and detection method and application thereof
  • Gastric cancer biomarker based on peripheral blood and detection method and application thereof
  • Gastric cancer biomarker based on peripheral blood and detection method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0052] Example 1: Collection and preservation of blood samples

[0053] Peripheral blood samples were collected from Jinling Hospital affiliated to Nanjing University School of Medicine, including 100 patients with gastric cancer, 30 patients with benign gastric disease, and 75 healthy subjects.

[0054] Take 0.5mL fresh EDTA-K2 anticoagulant blood, and immediately add 1.5-2mL RNALock blood RNA stabilizer (Tiangen Biochemical Technology (Beijing) Co., Ltd.).

[0055] Immediately cover the tube lid, mix upside down and mix 8-10 times, and leave it at room temperature for more than 2 hours. The mixed samples are stored in a low-temperature refrigerator at -20°C.

Embodiment 2

[0056] Example 2: RNA extraction from blood samples

[0057] Step 1: When purifying blood samples stored in blood RNA stabilizers, first place the samples at room temperature or in a 37°C water bath to raise them to room temperature. After centrifugation at 6600 rpm for 10 min, a pipette was used to aspirate the supernatant, and RNAprep Pure Blood Total RNA Extraction Kit (Tiangen Biochemical Technology (Beijing) Co., Ltd.) was used to extract total RNA from the blood. Refer to the kit instructions for specific operations.

[0058] Nanodrop2000 detects RNA concentration and purity, and performs follow-up operations.

Embodiment 3

[0059] Example 3: RNA m 6 A methylation quantitative detection

[0060] Use EpiQuik from Epigentek TM m 6 A RNA Methylation Quantification Kit (Colorimetric) kit, please refer to the kit instructions for specific operations.

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Abstract

The invention discloses a gastric cancer biomarker based on peripheral blood and a detection method and application thereof. The invention also discloses a reagent or a kit for diagnosing gastric cancer by detecting the RNA m6A level of peripheral blood. The specific peripheral blood RNA m6A provided by the invention can be used as an intermediate result in the diagnosis of gastric cancer by usingalone or in combination with related mRNA or in combination with a serum tumor marker detection combination, which is helpful for doctors to diagnose gastric cancer by combining clinical symptoms, medical history or other examination information.

Description

Technical field [0001] The invention belongs to the field of biotechnology, and particularly relates to a gastric cancer biomarker based on peripheral blood, and a detection method and application thereof. Background technique [0002] Gastric cancer, as a malignant tumor originating from the epithelium of the gastric mucosa, is one of the most common malignant tumors in the digestive system. In 2018, it is estimated that there will be more than 1 million new cases of gastric cancer worldwide, and about 783,000 deaths, ranking the fifth in the incidence of malignant tumors and the third in the fatality rate. More than 70% of new cases of gastric cancer are from developing countries, and nearly half of the cases are from East Asia. Therefore, the early diagnosis of gastric cancer is particularly important. At present, gastroscopy biopsy and detection of serum tumor-related markers are the main diagnostic tools. However, gastroscopy biopsy is very traumatic, and the commonly use...

Claims

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Application Information

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IPC IPC(8): C12Q1/6886C12N15/11
CPCC12Q1/6886C12Q2600/158
Inventor 汪俊军王红胜葛李晨
Owner NANJING UNIV
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