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Novel diagnostic method of acute myeloid leukemia and application thereof

A technology for acute myeloid and leukemia, which is applied in the field of tumor diagnosis and can solve problems such as difficult coverage

Pending Publication Date: 2020-09-11
锐甲生物技术徐州有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

First, for other types of tumor diagnostic molecular markers, such as point mutations, the mutation sites of the same gene are completely different in different tumors or even in different individuals of the same tumor, and it is difficult to fully cover them with existing technologies; while abnormal DNA methylation widely exists in Almost all types of tumors, and the abnormal areas of the same tumors are the same, so the detection is convenient and accurate

Method used

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  • Novel diagnostic method of acute myeloid leukemia and application thereof
  • Novel diagnostic method of acute myeloid leukemia and application thereof
  • Novel diagnostic method of acute myeloid leukemia and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0023] Example 1 By analyzing the DNA methylation data in TCGA and GEO databases, comparing the methylation of DNA methylation elements such as CYP26C1-DMR, DGKG-DMR, WT1-DMR, TRIM40-DMR in AML patients and normal people level difference

[0024] The present invention collects 450K DNA methylation chip data of 335 cases of AML patients and 38 cases of normal people from TCGA and GEO databases, and uses this data to analyze CYP26C1-DMR, DGKG-DMR, WT1-DMR, TRIM40-DMR, etc. 4 The methylation level of each methylation element is different in AML patients and normal people. In order to verify the correctness of this discovery, the inventors collected data from another DNA methylation detection method—genome-wide reduced methylation sequencing (RRBS), which included 143 AML patients and 10 normal individuals.

[0025] Results: Both DNA methylation microarray data and RRBS data showed that the methylation levels of four DNA methylation elements, including CYP26C1-DMR, DGKG-DMR, WT1-DM...

Embodiment 2

[0026] Example 2 Detection of DNA methylation levels of CYP26C1-DMR, DGKG-DMR, WT1-DMR, TRIM40-DMR and other elements

[0027] Target region DNA methylation sequencing is a method that uses high-throughput sequencing to detect DNA methylation levels in specific regions. It has the advantages of high detection accuracy, parallel detection of multiple sites, and low cost. In order to apply this method to detect the four DNA methylation elements found above, the inventors further narrowed the detection range of the methylation elements, and designed amplification primers for the narrowed methylation element regions (Table 2).

[0028] The specific steps of DNA methylation detection in the target region are as follows:

[0029] 1. Use Novozyme's blood DNA extraction kit (FastPure Blood DNA Isolation MiniKit) to extract peripheral blood DNA. The specific steps are as follows: Take 200 μl of peripheral blood, shake and mix. Add 400 μl of ACK LysisBuffer, mix well, centrifuge at 11...

Embodiment 3

[0035] Embodiment 3 utilizes logistic regression model to diagnose AML

[0036] Using the DNA methylation chip data, the inventors established a logistic regression model for diagnosing AML ( image 3 A), and validate the accuracy of the model using the validation dataset of DNA methylation microarray data. The specific calculation method is: bring the methylation rate of each DNA methylation element into the regression equation, and calculate the p value. If the p value calculated by the equation is greater than 0.5, it is judged as an AML patient, and if it is less than 0.5, it is judged as a normal person. Finally, the model is further validated by using the collected sequencing data of healthy people and AML patients.

[0037] Result: if image 3 As shown in B, the logistic regression model used in the verification data set has a prediction accuracy of 100% for normal people, 96.2% for AML patients, and an overall accuracy of 96.4%. The AUC curve shows that the predicti...

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Abstract

The invention discloses a group of acute myeloid leukemia specific DNA methylation elements CYP26C1-DMR, DGKG-DMR, WT1-DMR and TRIM40-DMR, and a method for assisting in diagnosing acute myeloid leukemia by detecting methylation levels of the elements. The group of DNA methylation elements is abnormally high in methylation in the acute myeloid leukemia, can be used as a specific tumor marker for the acute myeloid leukemia, and has the important clinical value.

Description

technical field [0001] The invention relates to the field of tumor diagnosis, in particular to the application of an epigenetic marker in the diagnosis of acute myeloid leukemia. Background technique [0002] The incidence of leukemia is increasing year by year in my country, and acute myeloid leukemia (AML) is the most common type of acute leukemia. Radiation exposure, exposure to chemicals, and even some chemotherapy procedures can trigger AML. AML cells derived from bone marrow and peripheral blood will overproliferate and hinder the production of normal blood cells, which will damage the patient's immune system and increase the risk of infectious diseases. At the same time, it is often accompanied by a series of symptoms such as shortness of breath, fatigue, and fever. The course of AML progresses very rapidly, and if left untreated, patients may die within months or even weeks. AML patients mainly come from the elderly population, and for the patient population over 6...

Claims

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Application Information

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IPC IPC(8): C12Q1/6886C12Q1/6858
CPCC12Q1/6886C12Q1/6858C12Q2600/154C12Q2600/16C12Q2523/125C12Q2535/122C12Q2531/113C12Q2537/143
Inventor 钱鹏旭姜蓬垒韩颖丽
Owner 锐甲生物技术徐州有限公司
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