Analysis method of circulating tumor DNA (deoxyribonucleic acid) at single-molecule level

An analysis method and single-molecule technology, which is applied in the fields of biochemistry and medical detection, can solve the problems of large sample usage and insufficient specificity, and achieve the effects of small sample volume, high detection sensitivity, and reduced error

Pending Publication Date: 2020-06-19
XUZHOU NORMAL UNIVERSITY
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

The problem with these methods is that the specificity is not high enough and the sample usage is large

Method used

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  • Analysis method of circulating tumor DNA (deoxyribonucleic acid) at single-molecule level
  • Analysis method of circulating tumor DNA (deoxyribonucleic acid) at single-molecule level
  • Analysis method of circulating tumor DNA (deoxyribonucleic acid) at single-molecule level

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experiment example

[0035] a. Add 2 μL of 8 μM carboxylated QD585 and QD655 stock solutions to 14 μL NaHCO 3 (50 mM, pH9.0) buffer solution, then added with cold NaHCO 3 (50 mM, pH 9.0) 4 μL of EDC (1 mg / mL), and incubate at room temperature in the dark for 25 min.

[0036] b. ssDNA1 (modified primary amino group at the 5' end) is immobilized on the surface of QD585 by reacting its primary amino group with the carboxyl group of QD585. Take 5.2 μL of ssDNA1 (12.2 μM) and 20 μL of the above-mentioned activated QD585 solution, mix well, and incubate at room temperature in the dark for 4 h. ssDNA3 (modified primary amino group at the 5' end) was immobilized on the surface of QD655 by reacting its primary amino group with the carboxyl group of QD655. Take 3.8 μL of ssDNA3 (16.9 μM) and 20 μL of the above-mentioned activated QD655 solution, mix well, and incubate at room temperature in the dark for 4 h.

[0037]c. Heat ssDNA1-QD585 and ssDNA2 in PBS (10 mM, pH 6.8, containing 300 mM NaCl) buffer sol...

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Abstract

The invention discloses an analysis method of circulating tumor DNA (deoxyribonucleic acid) at a single-molecule level, and provides an analysis method of circulating tumor DNA (ctDNA) at a single-molecule level. The analysis method takes a quantum dot dimer ratio as a quantitative basis, and avoids the condition that the intensity changes of light, electricity, magnetism, force and other signalsare used for carrying out quantitative analysis; errors caused by signal fluctuation are reduced, so that the concentration of the ctDNA in blood is rapidly, sensitively and accurately detected.

Description

technical field [0001] The invention relates to a method for analyzing circulating tumor DNA (ctDNA) at the single molecule level, which belongs to the technical field of biochemistry and medical detection. [0002] technical background [0003] Circulating tumor DNA (ctDNA) is a DNA fragment from the genome of tumor cells in human blood. Tumor cells in the lesions of tumor patients continuously metastasize, undergo apoptosis and necrosis in the human circulatory system, and release circulating tumor DNA into the blood. , and because it has the characteristics of tumor genome mutation, insertion, deletion, rearrangement, methylation, etc., and carries biological information such as tumor occurrence, development, metastasis, and recurrence, it is of great importance for the diagnosis, treatment, and prognosis evaluation of tumors. value. [0004] Tissue biopsy is currently the most routine tumor detection method in clinical diagnosis, but tissue biopsy has limitations such as...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6834
CPCC12Q1/6834C12Q2563/107C12Q2565/601C12Q2525/301
Inventor 盖宏伟武张健刘晓君
Owner XUZHOU NORMAL UNIVERSITY
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