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Mitochondrial DNA copy index variability detecting device

A detection device and mitochondrial technology, applied in the field of medical detection, can solve the problems of uncertain definition of the measurement unit of chrM, inability to compare and analyze, inconsistent results, etc., and achieve the effects of high stability, difficult misdiagnosis, and small random errors.

Active Publication Date: 2015-06-10
SHANGHAI MAJORBIO BIO PHARM TECH
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  • Summary
  • Abstract
  • Description
  • Claims
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AI Technical Summary

Problems solved by technology

The reported quantitative PCR technology for mitochondrial DNA has the following shortcomings: (1) only a section of chrM is intercepted as an indicator of chrM as a whole, and the random error is large; (2) it can only be used as an independent indicator, and cannot be compared with other DNA in the sample. (3) The method of model fitting is used for quantification, and there are many influencing factors, and the results produced by different models and different experimental procedures are inconsistent; (4) The definition of the measurement unit of chrM is uncertain

Method used

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Examples

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Embodiment 1

[0027] Example 1 A detection device for mitochondrial DNA copy number variability

[0028] This embodiment relates to the structural composition of a detection device for mitochondrial DNA copy number variability. The device includes a sample preparation unit, a sequencing unit, a calculation unit, and a comparison and judgment unit, wherein:

[0029] The sample preparation unit includes disposable venous blood collection needles, disposable vacuum blood collection tubes, blood anticoagulant EDTA, blood cell protection agent, and plasma free DNA purification kit. The sample preparation unit is used to draw peripheral blood from the vein of the subject and separate Get the cell-free DNA in its plasma.

[0030] The sequencing unit includes a quality inspection module, a high-throughput sequencing module, and a data optimization module, and the high-throughput sequencing module further includes a library construction kit and a high-throughput sequencing kit. The quality inspecti...

Embodiment 2

[0036] Embodiment 2 Determination method of the diagnostic limit value in the detection device of embodiment 1

[0037] In the detection device for mitochondrial DNA copy number variability provided in Example 1 of the present invention, the method for obtaining the diagnostic threshold value 3.046 preset by the comparison judgment unit:

[0038] (1) Get a group of (31) free DNA in the plasma of healthy people as a control sample group, carry out whole genome sequencing to these control samples with the same device and method as the sequencing unit in Example 1 of the present invention, each control sample Obtain at least 20M sequencing numbers, compare the sequencing results to the human reference genome, and calculate the relative ratio M of the mitochondrial DNA copy number and the nuclear DNA copy number of each control sample H and M H The natural logarithm of lnM H :

[0039] m H =n H (chrM) / len(chrM) / n H (chrN) / len(chrN),

[0040] where: n H (chrM) is the number...

Embodiment 3  Embodiment 1 test test example

[0059] This embodiment relates to a test example of cancer screening using the mitochondrial DNA copy number variability detection device of embodiment 1.

[0060] 1. Use the sample preparation unit in the detection device to draw 5ml of venous blood from 5 subjects, separate the plasma, and extract free DNA fragments from the plasma as samples to be tested:

[0061] Plasma separation method: Centrifuge 5ml of venous blood at 1000rpm / min for 10 minutes, take out the upper layer of plasma and then centrifuge at 8000rpm / min for 10 minutes, and absorb the upper layer of plasma for DNA extraction;

[0062] The free DNA fragments were extracted using: QIAamp DNA Blood Mini Kit (cat.no.51104), a DNA extraction kit from QIAGEN.

[0063] 2. Use the quality inspection module to perform quality inspection of the concentration and fragment size of the DNA fragments of the sample to be inspected:

[0064] Quality inspection standard: TBS380 is used to quantify the extracted DNA, and the ...

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Abstract

The invention relates to the technical field of medical detection, and discloses a mitochondrial DNA copy number variability detecting device. The device comprises a sequencing unit, a computing unit and a comparative judgment unit. The sequencing unit carries out whole genome sequencing on free DNA in a plasma sample of a person to be detected. The computing unit compares a sequencing result of the sample to be detected with a human reference genome and computes the mitochondrial DNA copy index of the sample to be detected. The comparative judgment unit compares the mitochondrial DNA copy index of the sample to be detected with a preset diagnosis boundary value, judges whether mitochondrial DNA copy number variability exists on the sample to be detected or not and can further judge whether the person to be detected has the cancered risk or not. According to the detecting device, mitochondrial DNA copy number abnormal conditions are recognized by combining the molecular biology sequencing technology and the biological information analysis technology and carrying out whole genome sequencing on the free DNA in blood, so that whether the person to be detected is cancered or not is judged, and early-stage noninvasive cancer screening is achieved.

Description

technical field [0001] The invention relates to the technical field of medical detection, in particular to a device capable of detecting the variability of mitochondrial DNA copy number and then applied to early diagnosis and screening of cancer. Background technique [0002] Cancer is a general term for malignant tumors. Cancer cells are characterized by unlimited and endless proliferation, which consumes a large amount of nutrients in the patient's body; cancer cells release a variety of toxins, causing a series of symptoms in the human body; cancer cells can also metastasize It grows and reproduces everywhere in the body, leading to emaciation, weakness, anemia, loss of appetite, fever, and eventually destroys the structure and function of tissues and organs, causing necrosis, hemorrhage and infection, and the patient eventually dies due to organ failure. [0003] At present, although the detection of gene mutation sites has been used to guide medication and as a standard...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12M1/34
Inventor 曾丰波杨功达陈昌岳韩继臣
Owner SHANGHAI MAJORBIO BIO PHARM TECH
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