3'-terminal-2'-O methylation modified miRNA marker combination related to gastric cancer diagnosis and application thereof

A marker and methylation technology, which is applied in the determination/test of microorganisms, biochemical equipment and methods, etc., can solve the problems of unsatisfactory clinical screening, high cost, and restrict the development of methylation detection, and achieve good application. Prospect, Sensitivity, Specificity, Effectiveness of Broad Health Screening

Pending Publication Date: 2020-10-30
NANJING UNIV
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  • Description
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  • Application Information

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Problems solved by technology

However, because most of the current methylation detection methods require mass spectrometry and sequencing, the operation is cumbersome, the co

Method used

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  • 3'-terminal-2'-O methylation modified miRNA marker combination related to gastric cancer diagnosis and application thereof
  • 3'-terminal-2'-O methylation modified miRNA marker combination related to gastric cancer diagnosis and application thereof
  • 3'-terminal-2'-O methylation modified miRNA marker combination related to gastric cancer diagnosis and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0039] Example 1 Screening of 2'-O methylated differential miRNAs at the specific plasma 3' end of patients with gastric cancer

[0040] (1) The subjects of the study were patients with gastric cancer and healthy controls. Plasma from patients with gastric cancer and healthy controls was obtained from Nanjing Eastern Theater General Hospital. The blood collection was approved by patients and healthy volunteers and approved by the Ethics Committee of Eastern Theater General Hospital. The sample consisted of 60 patients with primary gastric cancer and 60 normal healthy volunteers. All patients with gastric cancer were confirmed by biopsy and had not undergone surgery, chemotherapy, radiotherapy or other tumor-related treatments before blood collection. The sample age ranged from 41 to 69 years old, and there was no significant difference in age composition between the two groups (P=0.83). The experiment of the present invention consists of a test set (n=12) and a verification ...

Embodiment 2

[0048] Utilize the kit of the present invention to verify the sequencing results

[0049] Stem-loop tailing method to determine the degree of 2'-O methylation at the 3' end of miRNA Principle: The methylation modification at the 3'-end of miRNA can affect the tailing efficiency of the tailing method in the kit due to the existence of steric hindrance , so that the measured miRNA content decreases and the Ct value increases. However, the 2'-O methylation at the 3' end of the miRNA did not affect the miRNA content determined by the stem-loop method, and the Ct value remained unchanged. Therefore, two methods are used to detect the plasma content of miRNA, and the Ct values ​​measured by the two methods are subtracted, and the difference in Ct can reflect the difference in the degree of methylation in plasma miRNA, and explore the use of this method for gastric cancer. Screening diagnosis. The specific test method is:

[0050] (1) Stem-loop method to determine and screen the r...

Embodiment 3

[0071] Operating characteristic curve analysis (ROC curve analysis)

[0072] In order to clarify the diagnostic efficiency of the 6 miRNA methylation assays we screened in patients with gastric cancer, we performed ROC curve analysis on the experimental results. The results are shown in Table 3. The areas under the ROC curve of -2'-O methylation detection to distinguish gastric cancer patients from normal controls were: miR-451, 0.743; miR-1, 0.722; let-7f, 0.840; miR-92a, 0.679; miR- 10a, 0.685; miR-21, 0.691. The area under the curve of the combined detection of 6 miRNAs can reach 0.947, indicating that the combination of 6 miRNAs to detect the 3' end-2'-O methylation level by stem-loop tailing method can significantly screen patients with gastric cancer.

[0073] Table 3: ROC curve analysis of 6 miRNAs.

[0074]

[0075]

[0076] *The panel 1 Represents the joint detection of the above six indicators;

[0077] The panel 2 Represents the combined detection of miR-...

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Abstract

The invention discloses a 3'-terminal-2'-O methylation modified miRNA marker combination related to gastric cancer diagnosis and application of the 3 '-terminal-2'-O methylation modified miRNA markercombination. The 3 '-terminal-2'-O-methylation modified miRNA combination at least comprises miR-451, miR-1 and let-7f. The marker disclosed by the invention has the characteristics of economy, rapidness, sensitivity, good specificity and the like when being combined for clinical detection of gastric cancer, can be used for wide health screening, and has a good application prospect.

Description

technical field [0001] The invention belongs to the field of biological detection, and relates to a combination of 3'-2'-O methylation-modified miRNA markers related to the diagnosis of gastric cancer and an application thereof. Background technique [0002] Gastric cancer (GC) is a malignant tumor originating from gastric mucosal epithelium. It is recognized as the fifth most common cancer and the third leading cause of cancer death worldwide. Gastric cancer cases mostly occur in developing countries, with the highest incidence in Southeast Asia, among which the incidence of gastric cancer in China and Japan is the most prominent. The onset of gastric cancer is hidden, and most patients have already developed distant metastasis when symptoms appear, which seriously affects the prognosis of gastric cancer patients. The currently implemented screening methods for gastric cancer patients, such as endoscopy or traditional plasma tumor marker examination, are painful for patien...

Claims

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Application Information

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IPC IPC(8): C12Q1/6886
CPCC12Q1/6886C12Q2600/154C12Q2600/158C12Q2600/178
Inventor 陈熹韩嘉熠孙影吴雪娇杨紫琳
Owner NANJING UNIV
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