Unlock instant, AI-driven research and patent intelligence for your innovation.

A sedimentation aid for improving the extraction rate of free dna in plasma samples

A sedimentation aid and a technology in samples, applied in the direction of DNA preparation, recombinant DNA technology, etc., can solve the problems of low plasma circulating DNA content, insufficient free DNA yield, and inability to meet the detection of trace gene mutations, etc., to achieve Improve the extraction rate, easy to use, improve the effect of detection rate

Active Publication Date: 2022-05-06
麦凯(浙江)医疗器械有限公司 +1
View PDF0 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, even in patients with advanced tumors, the plasma circulating DNA (cfDNA) content of most patients is very low, which brings great challenges to the subsequent detection of ctDNA
The more commonly used plasma cfDNA extraction methods in current research usually include several steps of plasma separation from whole blood, lysing, column binding, washing and elution, and free nucleic acids can be extracted from plasma. The yield of cell-free DNA extracted by the plasma cell-free DNA extraction kit QIAamp Circulating Nucleic Acid Kit (QIAGEN) is still not high enough to meet the needs of trace gene mutation detection

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • A sedimentation aid for improving the extraction rate of free dna in plasma samples
  • A sedimentation aid for improving the extraction rate of free dna in plasma samples

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0022] A sedimentation aid for increasing the extraction rate of free DNA in plasma samples provided in this example is composed of the following formula, that is, added to each milliliter of plasma samples:

[0023] Human Cot-1 DNA 4.3mg;

[0024] Chelex-100 cation exchange resin 1.8mg.

[0025] 1) Take 10 cases of normal separated plasma samples, 4mL for each case; divide each case of plasma into 2 parts, 2mL respectively;

[0026] 2) Add the sedimentation aid of the present invention (namely: 8.6 mg of human Cot-1 DNA, 3.6 mg of Chelex-100 cation exchange resin) to one plasma sample of each case according to the above ratio, mix well and heat at 90°C 10min, then cooled down to room temperature;

[0027] 3) Cell-free DNA was extracted from two plasma samples at the same time. The extraction kit used the QIAamp Circulating Nucleic Acid Kit nucleic acid extraction reagent (product number: 55114) from Kaiser, Germany. The specific extraction operation was as follows:

[0028...

Embodiment 2

[0039] A sedimentation aid for increasing the extraction rate of free DNA in plasma samples provided in this example is composed of the following formula, that is, added to each milliliter of plasma samples:

[0040] Human Cot-1 DNA 4.5mg;

[0041] Chelex-100 cation exchange resin 2.0mg.

[0042] 1) Take plasma samples from 10 patients with EGFR gene mutation-positive non-small cell lung cancer, 4 mL for each case; divide each case of plasma into 2 parts, 2 mL each;

[0043] 2) Add the sedimentation aid of the present invention (i.e.: 9.0 mg of human Cot-1 DNA, 4.0 mg of Chelex-100 cation exchange resin) to one plasma sample of each case according to the above ratio, mix well and heat at 90°C 10min, then cooled down to room temperature;

[0044] 3) Cell-free DNA was extracted from two plasma samples at the same time. The extraction kit used the QIAamp Circulating Nucleic Acid Kit nucleic acid extraction reagent (product number: 55114) from Kaiser, Germany. The specific extra...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

PUM

No PUM Login to View More

Abstract

The invention discloses a sedimentation aid for improving the extraction rate of free DNA in plasma samples, which is composed of the following formula, that is, adding 3.2 to 5.3 mg of human Cot-1 DNA, Chelex-100 to each milliliter of plasma samples Cation exchange resin 1.4 ~ 2.2mg. Experiment shows: use the sedimentation aid provided by the present invention, can effectively remove interfering substance in blood plasma, can obviously improve the extraction yield of free DNA, and then can improve the detection rate of gene mutation in tumor free DNA (under the same extraction conditions , the extraction yield of free DNA can reach more than 99%, and then the detection rate of gene mutation in tumor free DNA can be increased from 70% to 90%); in addition, the sedimentation aid of the present invention is easy to use, only It needs to be added to the plasma sample and the existing extraction kit can be used to significantly improve the extraction rate of free DNA in the plasma sample.

Description

technical field [0001] The invention relates to a sedimentation aid for improving the extraction rate of free DNA in plasma samples, and belongs to the technical field of nucleic acid extraction. Background technique [0002] As a branch of in vitro diagnosis, liquid biopsy is to diagnose the genetic status of tumor patients by collecting blood from patients, and has broad application prospects in the field of clinical medicine. Among them, tumor cell-free DNA (ctDNA), as a non-invasive tumor marker, is considered to have a good development prospect. However, even in patients with advanced tumors, the plasma circulating DNA (cfDNA) content of most patients is very low, which brings great challenges to the subsequent detection of ctDNA. The more commonly used plasma cfDNA extraction methods in current research usually include several steps of plasma separation from whole blood, lysis, column binding, washing and elution, and free nucleic acids can be extracted from plasma, b...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

Application Information

Patent Timeline
no application Login to View More
Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/10
CPCC12N15/1003
Inventor 鹿亚超齐民强
Owner 麦凯(浙江)医疗器械有限公司