Kit for simultaneously extracting circulating tumor cell DNA and tumor free DNA

A technology of tumor cells and kits, which is applied in the field of kits for simultaneous extraction of circulating tumor cell DNA and tumor cell-free DNA, can solve problems such as poor stability of DNA content and affect the effectiveness of blood tests, and achieve easy mastery, low detection cost, The operation process is convenient and simple

Inactive Publication Date: 2017-03-08
武汉海吉力生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] With the development of the wide application of tumor molecular diagnosis, non-invasive detection of tumor mutation genes in peripheral blood has become a trend, but the current existing method is only a simple and rough replacement of the previ...

Method used

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  • Kit for simultaneously extracting circulating tumor cell DNA and tumor free DNA
  • Kit for simultaneously extracting circulating tumor cell DNA and tumor free DNA
  • Kit for simultaneously extracting circulating tumor cell DNA and tumor free DNA

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0028] Example 1: Extraction and analysis of mixed samples of free DNA and circulating tumor cell DNA in peripheral blood

[0029] In this example, tumor cell-free DNA and circulating tumor cell DNA were extracted from the peripheral blood of patients with non-small cell lung cancer, and the specific details are as follows.

[0030] 1. Sample processing

[0031] (1) 20 lung cancer patients were selected as the experimental subjects, and 5ml of venous blood was extracted from these 20 patients with EDTA anticoagulant tubes. In their FFPE tissue EGFR gene sequencing results (gold standard), 15 EGFR-positive patients and 5 EGFR-negative patients were used as sample donors, and the total positive rate was 75%. Mark and store at 4°C. Peripheral blood needs to be subjected to density gradient centrifugation within 3 hours.

[0032] (2) Dispense 3ml of Oncoquick separation solution into 15ml separation tubes (split into 20 tubes), and then gently add anticoagulated fresh periphera...

Embodiment 2

[0050] Example 2: Extraction and analysis of individual samples of circulating tumor cell DNA

[0051] The experimental procedure was the same as in Example 1, wherein the extracted samples were only the isolated CTC cells. This example demonstrates the enrichment capability of the kit for CTC cells and the separate extraction capability of CTC cell DNA. For test results, see Table 1-2 and Figure 4 .

Embodiment 3

[0052] Example 3: Extraction and analysis of a separate sample of peripheral blood cell-free DNA

[0053] The experimental procedure is the same as in Example 1, wherein the extracted sample is changed to the plasma separated in Example 1. This example demonstrates the separate extraction ability of the plasma free DNA of the kit. For test results, see Table 1-2 and Figure 5 .

[0054] Analysis of statistical results:

[0055] Statistical analysis was performed on the test results of the mixed samples of free DNA and circulating tumor cell DNA, the single sample of circulating tumor cell DNA, and the single sample of free DNA, and the results are as follows:

[0056] Table 1 Results Statistics

[0057]

[0058] Table 2. Data Analysis of Table 1 Results

[0059]

[0060] According to the above results, it can be seen that this kit can effectively separate and extract CTC DNA and free DNA, and can use the extracted DNA for related downstream detection. The positive ...

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Abstract

The invention discloses a kit for simultaneously extracting circulating tumor cell DNA and tumor free DNA. The kit comprises a cell density gradient separation medium, a porous diaphragm separation tube, a centrifugal tube with the volume of 5 mL and a blood free DNA extraction kit. By means of the kit, according to the characteristics that the mutant gene detection rate of the free DNA and the circulating tumor cell DNA has no relevance with occurrence, through the complementarity of detection results of the free DNA and the circulating tumor cell DNA, the two sources of the free DNA and the circulating tumor cell DNA are creatively superposed, the sample size is increased, thus, the detection rate of patient tumor mutant genes can be effectively increased, clinical detection requirements are met, non-invasive liquid biopsy is better and more widely applied, and more tumor patients of different condition types can obtain benefits from tumor molecular diagnosis. The use process is convenient and easy, the detection cost is far lower than that of the unicell detection technology, doctors can easily grasp the kit, and the kit is suitable for clinical application and popularization.

Description

technical field [0001] The invention relates to the technical field of tumor gene mutation detection, in particular to a method for qPCR tumor-related gene mutation detection by enriching and extracting circulating tumor cell DNA and tumor free DNA from human peripheral blood as templates, specifically a method for simultaneously extracting circulating tumor cell DNA and tumor free DNA as templates. Tumor cell-free DNA kit. Background technique [0002] With the rapid development of molecular biology, people's understanding of tumors has reached the molecular level. Many tumor-related genes have been discovered, and the targeted therapy of tumor drugs has achieved good results. According to the characteristics that the curative effect of different drugs is correlated with tumor genotype, detection of tumor genotype can help clinicians to adopt precise treatment plan. In May 2003, the U.S. Food and Drug Administration approved the small molecule gene target drug Gefitinib (...

Claims

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Application Information

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IPC IPC(8): C12N15/10
CPCC12N15/1003C12Q1/6806C12Q2523/32
Inventor 张志立黄俊郭骁赵平锋卢彦羽张全胡树军
Owner 武汉海吉力生物科技有限公司
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