Marker for early diagnosis, risk stratification and prognosis risk stratification of acute myelogenous leukemia and application thereof

A technique for acute myeloid, early diagnosis, applied in biochemical equipment and methods, microbial determination/examination, DNA/RNA fragments, etc.

Pending Publication Date: 2022-06-17
NINGBO FIRST HOSPITAL
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] Single-strand DNA-binding protein (SSBPs) family genes widely exist in nature, among which, the SSBP4 gene is located on chromosome 19p13.11, and there is no report on the relationship between SSBP4 gene and acute myeloid leukemia

Method used

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  • Marker for early diagnosis, risk stratification and prognosis risk stratification of acute myelogenous leukemia and application thereof
  • Marker for early diagnosis, risk stratification and prognosis risk stratification of acute myelogenous leukemia and application thereof
  • Marker for early diagnosis, risk stratification and prognosis risk stratification of acute myelogenous leukemia and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0033] Specific detection primers were designed for the SSBP4 gene, and the SSBP4 gene in cells was amplified by RT-qPCR and sequenced to verify the reliability of the specific detection primers.

[0034] The specific detection primers for the SSBP4 gene include:

[0035] Forward primer: 5'-TACGTTTATGAGTACCTGCTGCACATC-3' (SEQ ID No. 1);

[0036] Reverse primer: 5'-TTCTTCTCCCATCGGATCTCAGACAG-3' (SEQ ID No. 2);

[0037] At the same time, the specific detection primers for the internal reference gene GAPDH were designed, including:

[0038] Forward primer: 5'-ATGGGGAAGGTGAAGGTCG-3' (SEQ ID No. 3);

[0039] Reverse primer: 5'-GGGTCATTGATGGCAACAATATC-3' (SEQ ID No. 4).

[0040] The above two pairs of specific primers were synthesized by Shanghai Sangon Biological Company.

[0041] RT-qPCR amplification methods include:

[0042] (1) Cell collection and pretreatment

[0043] The MV4-11 cell line cultured in vitro was collected, washed twice with PBS, and 1 mL of RNAiso Plus rea...

Embodiment 2

[0059] The above-mentioned specific detection primers are used to detect and analyze the expression of SSBP4 gene in the bone marrow of patients with acute myeloid leukemia. The detection and analysis methods include:

[0060] (1) Collection and processing of bone marrow specimens

[0061] From June 2016 to August 2021, 148 new AML bone marrow samples and 40 normal control bone marrow samples were collected from Ningbo First Hospital. All participants signed the informed consent; bone marrow samples were collected from bone marrow cell culture flasks 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 clumps, repeated pipetting and mixing, and stored at -80°C until use.

[0062] (2) RNA extraction: same as Example 1;

[0063] (3) cDNA reverse transcription: same as Example 1;

[0064] (4) Quantitative PCR: th...

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Abstract

The invention discloses a marker for early diagnosis, risk stratification and prognosis risk stratification of acute myelogenous leukemia and application of the marker. A patentee finds that compared with a normal person, the expression quantity of SSBP4 in bone marrow of an acute myelogenous leukemia (hereinafter referred to as AML) patient is remarkably increased, meanwhile, the expression level of the SSBP4 gene is remarkably related to prognosis of the AML (non-APL) patient, and the AML (non-APL) patient with high SSBP4 expression has poorer overall survival and event-free survival, which prompts that the prognosis is poor; high expression of SSBP4 also prompts poor prognosis in a risk group (cytogenetics risk stratification) in AML, and the high expression of SSBP4 particularly plays a very important role in prompting early death of patients; the SSBP4 gene can be used as a marker for early diagnosis, risk stratification and prognosis risk stratification of acute myelogenous leukemia.

Description

technical field [0001] The invention belongs to the field of biotechnology, and particularly relates to a marker for early diagnosis, risk stratification and prognosis risk stratification of acute myeloid leukemia and its application. Background technique [0002] Acute myeloid leukemia (AML) is a highly heterogeneous hematopoietic malignancy characterized by differentiation arrest and clonal proliferation of hematopoietic stem and progenitor cells. Karyotype analysis is often used to evaluate the prognosis of AML patients. With the improvement of molecular biology technology, gene mutation and rearrangement have been widely used in clinical practice. Some molecular mutations, such as NPM1, FLT3-ITD, CEBPA, c-KIT, etc., were included in the NCCN (National Comprehensive Cancer Network) and ELN (European Leukemia Net) guidelines as markers for the AML risk stratification prognostic assessment system. [0003] With the continuous improvement of medical level, due to the discov...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6886C12N15/11
CPCC12Q1/6886C12Q2600/118C12Q2600/166C12Q2600/158
Inventor 陈莹牧启田牛婷婷邹多兵石淙吴莹张昭艺
Owner NINGBO FIRST HOSPITAL
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