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Reagent and kit for diagnosing acute change period of chronic myeloid leukemia based on LGALS3 splicing variant

A technology of variants and reagents, applied in the field of reagents and kits for diagnosis of CML blastic phase based on LGALS3 splicing variants, achieving high clinical application value, high sensitivity and specificity

Active Publication Date: 2021-03-19
THE SECOND AFFILIATED HOSPITAL TO NANCHANG UNIV
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  • Claims
  • Application Information

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Problems solved by technology

Although some previous research reports have confirmed that alternative splicing plays an important role in the development of CML, there is no report that LGALS3 alternative splicing may regulate the abnormal splicing of pre-mRNA and participate in the progression of CML. There is an urgent need for early, accurate and specific detection of

Method used

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  • Reagent and kit for diagnosing acute change period of chronic myeloid leukemia based on LGALS3 splicing variant
  • Reagent and kit for diagnosing acute change period of chronic myeloid leukemia based on LGALS3 splicing variant
  • Reagent and kit for diagnosing acute change period of chronic myeloid leukemia based on LGALS3 splicing variant

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Experimental program
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Embodiment 1

[0029] 1. Sample collection of CML patient group

[0030] According to the NCCN diagnostic criteria for CML in 2016, peripheral blood samples were collected from patients with chronic myeloid primary and blast disease who were not treated for the first time.

[0031] 2. Extraction of peripheral blood mononuclear cells

[0032] Ficoll density gradient centrifugation method: ①Put 5ml whole blood into a 50ml centrifuge tube, dilute with 5ml PBS solution, and mix gently; ②Take two 15ml centrifuge tubes and add 5ml Ficoll solution first. Then gently add the diluted blood to the upper layer of Ficoll with a pipette (5ml each) (to avoid mixing the two solutions); ③centrifuge at 2000rpm for 20min; ④use a pipette to draw the white cell layer (ie PBMC) into a clean 15ml ⑤Add PBS to 10-15ml, centrifuge at 1500rpm for 10min, remove the supernatant, add Trizol and mix well. Store at -80°C for later use.

[0033] 3. Total RNA extraction

[0034] TRIzol method: ① Add 1ml TRIzol / 5×106 PBM...

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Abstract

The invention discloses application of 3# intron retention and / or loss spliceosome LGALS3-R and / or LGALS3-M of LGALS3 in preparation of a reagent for diagnosing the acute change period of chronic myeloid leukemia or a kit containing the diagnostic reagent. The invention has relatively high sensitivity and specificity, and can observe the acute change period of chronic myeloid leukemia more simplyand accurately as soon as possible; therefore, treatment means can be adopted in time; and relatively high clinical application value is achieved.

Description

technical field [0001] The present invention relates to the application of the LGALS3 splicing variant as a marker in the diagnosis of patients with chronic myelogenous leukemia in blast phase, as well as the application in the preparation of corresponding detection reagents and kits. Background technique [0002] Chronic myeloid leukemia (CML) is a malignant proliferative disease originating from pluripotent hematopoietic stem cells, accounting for about 20% of human leukemias. Studies have found that about 95% of CML patients have abnormal cytogenetic changes, that is, reciprocal translocation of chromosome 9 and chromosome 22 to form Philadelphia chromosome (philadelphia, Ph) and Bcr-Abl fusion gene. BCR / ABL fusion protein has strong tyrosine kinase activity, abnormally activates multiple downstream signaling pathways, and stimulates abnormal proliferation and malignant transformation of hematopoietic cells. The course of CML is usually divided into three stages: chronic...

Claims

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Application Information

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IPC IPC(8): C12Q1/6886
CPCC12Q1/6886C12Q2600/112C12Q2600/156C12Q2600/158
Inventor 黄波赵翠李书琪章海斌刘静王小中
Owner THE SECOND AFFILIATED HOSPITAL TO NANCHANG UNIV
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