Looking for breakthrough ideas for innovation challenges? Try Patsnap Eureka!

Exosome RNA for colorectal adenoma diagnosis and application thereof

A colorectal and exosome technology, applied in the direction of DNA / RNA fragments, recombinant DNA technology, microbial measurement / inspection, etc., can solve problems such as limitations, missed diagnosis, and low risk

Active Publication Date: 2020-08-04
BEIJING FRIENDSHIP HOSPITAL CAPITAL MEDICAL UNIV
View PDF1 Cites 2 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, colorectal adenomas are usually small in diameter and often located in the colonic mucosal folds, resulting in missed diagnosis, and the detection rate of lesions under colonoscopy depends on the ability and experience of the examiner, the configuration of the colonoscope, the degree of cleanliness of the subject's bowel, There are many factors such as the degree of cooperation of the subjects, and there is a certain probability of serious complications such as bleeding and perforation. There are certain restrictions on performing this test in all groups of people.
The current non-invasive detection methods in clinical practice include fecal occult blood detection, blood tumor marker detection, etc. Although the detection method is simple, the subject has a high degree of acceptance, and the risk is low, there are still some deficiencies in the sensitivity and specificity of lesion detection , so it is particularly important to develop a highly sensitive and specific non-invasive detection method for colorectal adenoma

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Exosome RNA for colorectal adenoma diagnosis and application thereof
  • Exosome RNA for colorectal adenoma diagnosis and application thereof
  • Exosome RNA for colorectal adenoma diagnosis and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0046] Example 1 Screening of exosomal RNA molecular markers related to colorectal adenomas based on high-throughput sequencing

[0047]The blood from 15 cases of early colorectal adenoma patients and 15 cases of healthy subjects in the control group was not less than 10mL, and the plasma was separated by classical ultracentrifugation (Evaluation of circulating small extracellularvesicles derived miRNAs as biomarkers 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) isolated exosomes in plasma , QIAGEN miRNeasy mini kit was used to extract RNA, and the prepared RNA was subjected to trace chain-specific long RNA sequencing (Evaluation of circulating small extracellular vesicles derived miRNAs as biomarkers 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 extr...

Embodiment 2

[0050] Example 2 mRNA and lncRNA detection system based on fluorescence quantitative PCR platform

[0051] 1. Reverse transcription system

[0052] Using TAKARA's PrimeScript TM RT reagent Kit (Perfect Real Time) and PremixEx Taq TM (Probe qPCR) kit for reverse transcription and qPCR detection.

[0053] Prepare the reverse transcription reaction system according to the components listed in Table 2 (the reaction solution is prepared on ice), and then put it into the PCR machine for the reaction. The reaction conditions are 37°C for 60min, 85°C for 5s, 12°C∞, and the reverse transcription is completed. After adding 50 μL DEPC H 2 O dilution, take 3uL as template, carry out PCR reaction.

[0054] Table 2 Reverse transcription reaction system

[0055]

[0056]

[0057] 2. qPCR system

[0058] The primers and probes for detecting exosomal miRNA molecular markers include:

[0059] Primers and probes for detecting lnc-MKRN2-42:1: the upstream primer of the lnc-MKRN2-42:1...

Embodiment 3

[0071] Example 3 Using RNA molecular markers for early diagnosis and detection of colorectal adenomas

[0072] 1) Sample collection

[0073] Collect 10 mL of blood including hospital-diagnosed early (stage I and II) colorectal adenoma patients (24 cases), colorectal benign patients and healthy controls (53 cases in total), and separate plasma.

[0074] 2)Exosome RNA extraction

[0075] Plasma exosomes were isolated using Qiagen's commercial ExoEasy kit or Enzekangtai's Exosupur, and the isolated exosomes were extracted with Qiagen miReasy mini kit to extract RNA from exosomes, and Agilent 2100 was used to detect RNA concentration and mass, and record the RNA concentration.

[0076] 3) RNA two-step detection system

[0077] Using the mRNA and lncRNA detection system based on the fluorescence quantitative PCR platform in Example 2, the plasma exosomal RNA extracted in step 2) was detected, and the Ct value of the target RNA was detected.

[0078] 4) Result:

[0079] Plasma ...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

PUM

No PUM Login to View More

Abstract

The invention relates to the field of biological detection, in particular to biological molecular detection, and more particularly relates to a group of exosome RNA molecular markers related to colorectal adenoma and application thereof.

Description

technical field [0001] The invention relates to the field of biological detection, more particularly to biological molecular detection, and more particularly to a group of exosome RNA molecular markers related to colorectal adenomas and applications thereof. Background technique [0002] Colorectal adenomas originate from the colorectal mucosal glandular epithelium and belong to tumor polyps from histological point of view. The canceration rate is reported in the literature to be 2.9%-9.4%, especially for individual pathological types such as villous tubular adenomas. It has been reported that most colorectal cancers develop from colorectal adenomas, so it is generally believed that colorectal adenomas are precancerous lesions of colorectal cancer and are closely related to the occurrence and development of colorectal cancer. In my country, epidemiology shows that the incidence and mortality of colorectal cancer are increasing year by year. Timely detection and treatment of ...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

Application Information

Patent Timeline
no application Login to View More
IPC IPC(8): C12Q1/6886C12N15/113
CPCC12Q1/6886C12Q2600/118C12Q2600/158C12Q2600/178
Inventor 张澍田朱圣韬闵力孔关义赵立波
Owner BEIJING FRIENDSHIP HOSPITAL CAPITAL MEDICAL UNIV
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Patsnap Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Patsnap Eureka Blog
Learn More
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic, Popular Technical Reports.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com