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Exosome RNA molecular marker combination used for early diagnosis of colorectal cancer and application of exosome RNA molecular marker combination used for early diagnosis of colorectal cancer

A colorectal cancer, molecular marker technology, applied in the field of biological detection, can solve the problems of low specificity and sensitivity, limited clinical application, low sensitivity, etc.

Active Publication Date: 2020-07-28
BEIJING FRIENDSHIP HOSPITAL CAPITAL MEDICAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, fecal occult blood test (FOBT) and fecal DNA test (FDT) are two feasible non-invasive screening methods in clinical practice. Although both methods can help reduce the mortality of colorectal cancer, their specificity is low. and sensitivity have limited its clinical application
Blood septin9 methylation detection is a non-invasive detection method based on blood, but its detection sensitivity is not high

Method used

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  • Exosome RNA molecular marker combination used for early diagnosis of colorectal cancer and application of exosome RNA molecular marker combination used for early diagnosis of colorectal cancer
  • Exosome RNA molecular marker combination used for early diagnosis of colorectal cancer and application of exosome RNA molecular marker combination used for early diagnosis of colorectal cancer
  • Exosome RNA molecular marker combination used for early diagnosis of colorectal cancer and application of exosome RNA molecular marker combination used for early diagnosis of colorectal cancer

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0043] Example 1 Screening of Exosomal RNA Molecular Markers Related to Colorectal Cancer Based on High-throughput Sequencing

[0044]No less than 10 mL of blood was collected from 15 patients with early colorectal cancer and 15 healthy subjects in the control group. comparison with plasma total miRNAs, L Min, S Zhu, L Chen, X Liu, R Wei, L Zhao, Y Yang, Z Zhang, G Kong, Journal of extracellular vesicles 8(1), 1643670) isolated exosomes in plasma, RNA was extracted using QIAGEN miRNeasy mini kit, and the prepared RNA was subjected to microstrand-specific RNA library construction and sequencing (Evaluation of circulating small extracellular vesicles derived miRNAs asbiomarkers of early colon cancer: a comparison with plasma total miRNAs, L Min, S Zhu, L Chen, X Liu, R Wei, L Zhao, Y Yang, Z Zhang, G Kong, Journal of extracellular vesicles 8(1), 1643670). Bioinformatics analysis was carried out on the sequencing results, and the differentially expressed RNAs in patients with ea...

Embodiment 2

[0047] Example 2 miRNA, mRNA and lncRNA detection system based on fluorescent quantitative PCR platform

[0048] 1. miRNA detection system

[0049] 1) miRNA reverse transcription reaction system

[0050] miRNA reverse transcription reagents, enzymes and oligdT were purchased from TAKARA, standard products were synthesized from Shanghai Yingwei Jieji, and specific reverse transcription primers were synthesized by Suzhou Synbio. A 20 μL reverse transcription system was used, as shown in Table 2.

[0051] Table 2 miRNA reverse transcription system

[0052]

[0053] 2) qPCR reaction system:

[0054] The PCR reaction mixture was purchased from TAKARA, the upstream primers and probes, namely the universal downstream primers, were synthesized by Suzhou Synbio, and the fluorescent quantitative PCR instrument was ABI 7500. The PCR reaction system is shown in Table 3:

[0055] Table 3 miRNA qPCR reaction system

[0056]

[0057] 2. mRNA and lncRNA detection system

[0058] ...

Embodiment 3

[0080] Example 3 Early Diagnosis of Colorectal Cancer Using RNA Molecular Markers

[0081] 1) Sample collection

[0082] Collect 10 mL of blood from a total of 100 subjects, including hospital-diagnosed early (stage I and stage II) colorectal cancer patients (47 cases), benign colorectal patients and healthy control samples (53 cases in total), and separate plasma.

[0083] 2) Exosomal RNA extraction

[0084] Plasma exosomes were separated using Qiagen’s commercial ExoEasy kit or Enze Kangtai’s Exosupur, and the isolated exosomes were extracted from exosomes with Qiagen miReasy mini kit, and the RNA concentration and mass, and record the RNA concentration.

[0085] 3) RNA detection system

[0086] Using the mRNA and lncRNA detection system based on the fluorescent quantitative PCR platform in Example 2, detect the plasma exosomal RNA extracted in step 2), detect the Ct value of the target RNA, and calculate the relative expression level according to the Ct value and the rel...

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Abstract

The invention relates to the field of biological detection, in particular to biological molecular detection and a combination of exosome RNA markers related to colorectal cancer and application of thecombination of the exosome RNA markers related to colorectal cancer.

Description

technical field [0001] The present invention relates to the field of biological detection, more specifically to biological molecular detection, and more specifically to a group of exosomal RNA molecular marker combinations and applications related to colorectal cancer. Background technique [0002] Colorectal cancer (CRC) is one of the common malignant tumors of the digestive system. In my country, its morbidity and mortality are second only to gastric cancer, esophageal cancer and primary liver cancer. Colorectal cancer has almost no obvious symptoms in the early stage of the disease. As the lesion gradually increases, symptoms such as diarrhea, constipation, abdominal pain and even blood in the stool will appear, and in the late stage, systemic symptoms such as anemia and weight loss will appear. In recent years, the incidence and mortality of colorectal cancer in my country have been on the rise. Statistics show that the incidence and mortality of colorectal cancer in my...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6886C12N15/113
CPCC12Q1/6886C12Q2600/118C12Q2600/178C12Q2600/106
Inventor 朱圣韬闵力孔关义刘翔张澍田
Owner BEIJING FRIENDSHIP HOSPITAL CAPITAL MEDICAL UNIV
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