Enhancer for free circulating tumor DNA extraction, kit and method for extracting free circulating tumor DNA in peripheral blood

A technology of enhancer and kit, which is applied in the field of biomedicine, can solve the problems of general extraction efficiency, difficulty in detection, and large difference, and achieve the effects of broadening the application range, increasing extraction yield, and improving extraction efficiency

Active Publication Date: 2019-02-22
BEIJING USCI MEDICAL DEVICES CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, there are still some technical challenges in using ctDNA to detect gene mutations, mainly as follows: 1) ctDNA content varies from person to person, and most people have low content; 2) ctDNA fragments are relatively small, most of them are about 180bp, Fragments are distributed between 100bp and 400bp; 3) The proportion of tumor-associated DNA in ctDNA varies greatly from person to person, and is often difficult to detect due to the small proportion
These limit the wide application of ctDNA in tumor detection. Therefore, efficient and convenient ctDNA extraction is an important factor affecting the wide application of ctDNA in tumor detection.
[0004] At present, commercialized ctDNA extraction kits mainly include column extraction method and magnetic bead method. It is simple and fast, does not require complex instruments, and can be combined with extraction workstations to reduce manual operations. However, the extraction efficiency of commercial magnetic bead extraction kits is average at present, and the problem of extraction efficiency needs to be solved urgently

Method used

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  • Enhancer for free circulating tumor DNA extraction, kit and method for extracting free circulating tumor DNA in peripheral blood
  • Enhancer for free circulating tumor DNA extraction, kit and method for extracting free circulating tumor DNA in peripheral blood
  • Enhancer for free circulating tumor DNA extraction, kit and method for extracting free circulating tumor DNA in peripheral blood

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Embodiment 1

[0062] The kit of this embodiment consists of the following reagents:

[0063] 1. Magnetic bead suspension G was purchased from Life Company;

[0064] 2. SDS solution was purchased from Life Company with a concentration of 10%;

[0065] 3. Lysis binding solution: 45mmol / L Tris-HCl, 120mmol / LNaCl, 30mmol / L disodium edetate, 10mol / L guanidine isothiocyanate, 2mol / L potassium acetate, 5wt% Tween 20, pH The value is 7.6;

[0066] 4. Proteinase K was purchased from Life Company;

[0067] 5. Protease dissolving buffer: 45mmol / L Tris-HCl, 120mmol / LNaCl;

[0068] 6. Washing liquid 1: 45mmol / L Tris-HCl, 120mmol / LNaCl, 30mmol / L EDTA disodium, 1.5wt% Triton, pH value is 8.0;

[0069] 7. Washing solution 2: 80% ethanol;

[0070] 8. Eluent AE: nuclease-free water;

[0071] 9. Enhancer: the final concentration is 2.5 μg / μL Carrier RNA, 2.5 μg / μL polyadenylic acid, 2.5 μg / μL glycogen, 2.5 μg / μL linear acrylamide (linear acrylamide), 2.5 μg / μL yeast tRNA, 2.5 μg / μL salmon sperm DNA, 2....

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Abstract

The invention discloses an enhancer for free circulating tumor DNA extraction, a kit and a method for extracting free circulating tumor DNA in peripheral blood, wherein the enhancer comprises 1-5 [mu]g / [mu]L of carrier RNA, 1-5 [mu]g / [mu]L of polyadenylic acid, 1-5 [mu]g / [mu]L of glycogen, 0-5 [mu]g / [mu]L of linear acrylamide, 1-5 [mu]g / [mu]L of yeast tRNA, 2-10 [mu]g / [mu]L of salmon sperm DNA and / or menhaden sperm DNA and water. The enhancer for the free circulating tumor DNA (ctDNA) extraction provided by the invention can be used in combination with a conventional lysed binding solution, awashing buffer, an eluant and the like, can be specifically bound to ctDNA and deposited, and is capable of significantly improving the extraction efficiency and the extraction yield of ctDNA.

Description

technical field [0001] The invention relates to the technical field of biomedicine, in particular to an enhancer for extracting free circulating tumor DNA, a kit and a method for extracting free circulating tumor DNA in peripheral blood. Background technique [0002] With increasing morbidity and mortality, cancer has become the leading cause of death and a major public health problem in China. Cancer treatment methods are diverse, including radiotherapy, chemotherapy, and emerging targeted therapy and immunotherapy; among them, targeted therapy is an important component of precision medicine, and it is particularly important for the screening of targeted gene-specific populations. Cancer genetic testing It is directly related to the therapeutic efficiency of drugs for patients. At present, the main sample types used for detection are tumor tissue or puncture samples; however, tissue samples are invasive operations, which are sometimes difficult to implement, and these oper...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/10
CPCC12N15/1013
Inventor 潘晓西姬晓勇霍旭王建伟伍启熹刘倩刘珂弟唐宇
Owner BEIJING USCI MEDICAL DEVICES CO LTD
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