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Marker for auxiliary diagnosis, prognosis diagnosis or risk stratification of acute myeloid leukemia and application thereof

An acute myeloid and auxiliary diagnosis technology, applied in the direction of microbial determination/testing, biochemical equipment and methods, DNA/RNA fragments, etc., can solve problems such as incompleteness

Pending Publication Date: 2021-11-12
NINGBO FIRST HOSPITAL
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, cytogenetic-based risk stratification of AML is imperfect because most patients are at intermediate cytogenetic risk

Method used

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  • Marker for auxiliary diagnosis, prognosis diagnosis or risk stratification of acute myeloid leukemia and application thereof
  • Marker for auxiliary diagnosis, prognosis diagnosis or risk stratification of acute myeloid leukemia and application thereof
  • Marker for auxiliary diagnosis, prognosis diagnosis or risk stratification of acute myeloid leukemia and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0035] Specific detection primers were designed for the ARF3 gene, and the ARF3 gene in cells was amplified by RT-QPCR and sequenced to verify the reliability of the specific detection primers.

[0036] The specific detection primers of the ARF3 gene include:

[0037] Forward primer: 5'-GGAAGAGCTGATGAGAATGCTGG-3' (SEQ ID No.1);

[0038] Reverse primer: 5'-ACCAGTTACGGTGACGAAGGGA-3' (SEQ ID No.2);

[0039] The specific detection primers for the internal reference gene GAPDH were designed together, including:

[0040] Forward primer: 5'-ATGGGGAAGGTGAAGGTCG-3' (SEQ ID No.3);

[0041] Reverse primer: 5'-GGGTCATTGATGGCAACAATATC-3' (SEQ ID No. 4).

[0042] Both pairs of specific primers were synthesized by Qingke Biological Co., Ltd. (Hangzhou).

[0043] RT-QPCR amplification methods include:

[0044] (1) Cell collection and pretreatment

[0045]The THP-1 cells cultured in vitro were collected, washed twice with PBS, and 1 mL of RNAisoPlus reagent (purchased from TaKaRa) was ad...

Embodiment 2

[0061] The above-mentioned specific detection primers were used to detect and analyze the expression of the ARF3 gene in the bone marrow of patients with acute myeloid leukemia. The detection and analysis methods include:

[0062] (1) Collection and processing of bone marrow samples

[0063] A total of 116 AML bone marrow samples and 15 normal control bone marrow samples were collected from Ningbo First Hospital from June 2016 to June 2020. All participants signed the informed consent; bone marrow samples were collected using bone marrow cell culture bottles and recorded Patient information: mononuclear cells were collected by density gradient centrifugation of lymphocyte separation medium (Ficoll), and finally RNAiso Plus reagent was added to the collected mononuclear cell mass, mixed by repeated pipetting, and stored at -80°C until use.

[0064] (2) RNA extraction: same as Example 1;

[0065] (3) cDNA reverse transcription: same as Example 1;

[0066] (4) Quantitative PCR:...

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Abstract

The invention discloses a marker for auxiliary diagnosis, prognosis diagnosis or risk stratification of acute myeloid leukemia and application thereof. After multiple experiments, the patentee finds that compared with a normal person, the expression quantity of ARF3 in bone marrow of an acute myeloid leukemia (hereinafter referred to as AML) patient is remarkably increased, which indicates that the ARF3 gene can be used as an auxiliary diagnosis marker of AML; meanwhile, the patentee also finds that the expression level of the ARF3 gene is significantly related to the prognosis of the AML patient, and the AML patient with high ARF3 expression has poorer overall survival and event-free survival, which prompts that the prognosis is poor, especially in a risk group in cytogenetics risk stratification; and therefore, the ARF3 gene can be used as a marker for prognostic diagnosis or risk stratification of AML.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a marker for auxiliary diagnosis, prognosis diagnosis or risk stratification of acute myeloid leukemia and its application. Background technique [0002] Acute myeloid leukemia (AML) is a malignant clonal disease with high heterogeneity in cell morphology, molecular biology, cytogenetics, immunophenotype, and clinical manifestations. In recent years, although the complete remission rate and 5-year disease-free survival rate of AML patients have improved compared with the previous ones; Therefore, it is increasingly urgent to find new therapeutic targets and prognostic biomarkers. [0003] Altered expression or mutated forms of various genes involved in leukemia cell proliferation or survival are not only related to disease pathogenesis, but also important in predicting treatment response and survival. Although mutations in genes such as NPM1, FLT3, C-kit, C / EBPA, and RU...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6886C12N15/11
CPCC12Q1/6886C12Q2600/158C12Q2600/118
Inventor 邹多兵牧启田陈莹欧阳桂芳邬宁宁张怡
Owner NINGBO FIRST HOSPITAL
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