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Exosomal RNA for the diagnosis of colorectal adenoma and its application

A colorectal and adenoma technology, applied in the field of biological detection, can solve the problems of limitation, missed diagnosis, low risk, etc., and achieve high sensitivity and specificity, and good patient compliance.

Active Publication Date: 2021-05-18
BEIJING FRIENDSHIP HOSPITAL CAPITAL MEDICAL UNIV
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  • 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

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  • Exosomal RNA for the diagnosis of colorectal adenoma and its application
  • Exosomal RNA for the diagnosis of colorectal adenoma and its application
  • Exosomal RNA for the diagnosis of colorectal adenoma and its application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0046] Example 1 Screening of exosomal RNA molecular markers associated with colorectal adenoma based on high-throughput sequencing

[0047]No less than 10 mL of blood was collected from 15 patients with early colorectal adenoma and 15 healthy subjects in the control group. a comparison with plasma total miRNAs, L Min, S Zhu, L Chen, X Liu, R Wei, L Zhao, Y Yang, Z Zhang, GKong, 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 trace-specific long 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 long-chain RNAs with differential express...

Embodiment 2

[0050] Embodiment 2 mRNA and lncRNA detection system based on fluorescent 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 reaction. 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 exosome miRNA molecular markers include:

[0059] Primers and probes for detecting lnc-MKRN2-42:1: the upstream primer of the lnc-MKRN2-42:1 is specifically the nucleotide sequence shown in SEQ ID NO:1, and the downstream primer is specifically as show...

Embodiment 3

[0071] Example 3 Early diagnosis and detection of colorectal adenoma using RNA molecular markers

[0072] 1) Sample collection

[0073] Collect 10 mL of blood from patients with early (stage I and stage II) colorectal adenomas (24 cases), benign colorectal patients and healthy controls (53 cases in total) diagnosed by the hospital, and separate plasma.

[0074] 2) Exosomal RNA extraction

[0075] 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.

[0076] 3) RNA two-step detection system

[0077] The mRNA and lncRNA detection system based on the fluorescent quantitative PCR platform in Example 2 was used to detect the plasma exosomal RNA extracted in step 2), and to detect the Ct value of the target RNA.

[0078] 4) Results:

[0079] The Ct values ​​of lnc-MKRN2-42:1, PPDPF,...

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Abstract

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.

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 exosome RNA molecular markers related to colorectal adenoma and applications thereof. Background technique [0002] Colorectal adenoma originates from the glandular epithelial tissue of the colorectal mucosa, and is a neoplastic polyp from the histological point of view. It has been reported in the literature that its cancerous rate is 2.9%-9.4%, especially for individual pathological types such as villous tubular adenoma. It has been reported that most colorectal cancers develop from colorectal adenomas. Therefore, 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 ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
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
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