Screening method and application of aptamer for detecting acute promyelocyte leukemia

A technology for promyelocytic and leukemia, which is applied in biochemical equipment and methods, measuring devices, determination/examination of microorganisms, etc., can solve the problems that the specificity and accuracy of leukemia diagnosis needs to be further improved.

Inactive Publication Date: 2017-10-10
BEIHUA UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] In summary, there is currently a lack of comprehensive understanding of the mechanism of the occurrence and development of leukemia in clinical practice, and the specificity and accuracy of leukemia diagnosis need to be further improved

Method used

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  • Screening method and application of aptamer for detecting acute promyelocyte leukemia
  • Screening method and application of aptamer for detecting acute promyelocyte leukemia
  • Screening method and application of aptamer for detecting acute promyelocyte leukemia

Examples

Experimental program
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Effect test

Embodiment 1

[0056] 1. Synthesis of random single-stranded DNA library and primers

[0057] Random ssDNA library: 5'-ATC CAG AGT GAC GCA GCA-40nt-TGG ACA CGG TGG CTT

[0058] AGT-3'

[0059] Upstream fluorescein-labeled primer: 5'-FITC-ATC CAG AGT GAC GCA GCA-3'

[0060] Downstream biotinylated primer: 5'-BIO-ACT AAG CCA CCG TGT CCA-3'

[0061] Upstream primer: 5'-ATC CAG AGT GAC GCA GCA-3'

[0062] Downstream primer: 5'-ACT AAG CCA CCG TGT CCA-3'

[0063] 2. Cell-SELEX screening process

[0064] Cell-SELEX screening to obtain nucleic acid aptamers that specifically recognize acute promyelocytic leukemia

[0065] 2.1 Dissolve 1 μl of the 10 μM random ssDNA library above in 350 μl binding buffer, place in a constant temperature water bath at 95°C for 5 minutes, and then place it on an ice box immediately for use.

[0066] 2.2 Add 10% newborn bovine serum to the 1640 culture medium to cultivate acute promyelocytic leukemia HL-60 cells. When the number of cells reaches 5×10 6 / ml was t...

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Abstract

The invention provides an aptamer specifically combining acute promyelocyte leukemia HL-60 cells, and further provides a screening method and a clinical application of the aptamer. According to the screening method, six types of aptamers with different sequences are screened, and combining capacity among the aptamers and the acute promyelocyte leukemia HL-60 cells is different. The screening method includes the steps: synthesizing random single-strand DNA (deoxyribonucleic acid) libraries and primers; performing Cell-SELEX screening; performing library amplification; preparing a DNA single-strand library; repeating multiple screening and subduction screening; detecting stability after screening; detecting specificity of the aptamers.

Description

technical field [0001] The invention relates to the research field of acute promyelocytic leukemia detection technology, more specifically, it relates to a nucleic acid aptamer that can be used to detect acute promyelocytic leukemia at the molecular level and its screening method and application. Background technique [0002] Leukemia is a relatively common malignant tumor of the hematopoietic system, characterized by abnormal proliferation of leukemia cells in the bone marrow and other hematopoietic tissues, infiltration and destruction of various organs and tissues, resulting in structural and functional damage to the tissues and organs. The disease has a high mortality rate, great harm, and the disease progresses rapidly. If it is not treated in time, the patient usually dies within weeks or months after the disease. Currently, the diagnosis and treatment methods for acute leukemia, including acute promyelocytic leukemia (APL), are very limited. APL is a special type of ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/115C12N15/10G01N33/574
CPCC12N15/1048C12N2310/16G01N33/57426C12Q2525/205C12Q2563/143C12Q2563/149
Inventor 高丽君杜冰洋王诗洋王昶夏薇于丹孙娜张亚丽高爽刘佳张迪郝梦哲王梦主
Owner BEIHUA UNIV
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