Reagent for diagnosing chronic myeloid leukemia blast crisis based on hnRNPA1 splicing variant and kit

A technology of variants and reagents, applied in the field of reagents and kits for diagnosing CML blastic phase based on hnRNPA1 splicing variants, can solve problems such as lack of alternative splicing, and achieve high clinical application value, high sensitivity and specificity Effect

Active Publication Date: 2019-06-14
THE SECOND AFFILIATED HOSPITAL TO NANCHANG UNIV
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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 on alternative splicing, including the report that hnRNPA1 alte

Method used

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  • Reagent for diagnosing chronic myeloid leukemia blast crisis based on hnRNPA1 splicing variant and kit
  • Reagent for diagnosing chronic myeloid leukemia blast crisis based on hnRNPA1 splicing variant and kit
  • Reagent for diagnosing chronic myeloid leukemia blast crisis based on hnRNPA1 splicing variant and kit

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Embodiment 1

[0032] 1. Sample collection of CML patients and healthy controls

[0033] 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, as well as peripheral blood samples from the healthy physical examination group.

[0034] 2. Extraction of peripheral blood mononuclear cells

[0035] 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...

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Abstract

The invention discloses application of No.8 exon skip/ retained spliceosome (hnRNPA1-a and hnRNPA1-b) of hnRNPA1 in preparing a reagent for diagnosing chronic myeloid leukemia blast crisis or in a kitcontaining the diagnostic reagent. The reagent has higher sensitivity and specificity, can observe the chronic myeloid leukemia blast crisis more simply, accurately and early so as to timely take treatment means, and has higher clinical application value.

Description

technical field [0001] The present invention relates to the application of hnRNPA1 splicing variant as a marker in diagnosing patients with chronic myelogenous leukemia in blast phase, as well as the application in preparing 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 phase, accelerate...

Claims

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Application Information

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IPC IPC(8): C12Q1/6886G01N33/574
Inventor 王小中李书琪刘静张静黄波
Owner THE SECOND AFFILIATED HOSPITAL TO NANCHANG UNIV
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