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Kit, device and method for detecting copy number of embryo chromosomes

A chromosome copy number and kit technology, which is applied in biochemical equipment and methods, and the determination/inspection of microorganisms, can solve problems such as inappropriate detection of changes in chromosome copy number, and achieve accurate chromosome number or chromosome local copy number and sequence. The effect of reducing the number and reducing the cost of sequencing

Active Publication Date: 2011-06-29
HANGZHOU BERRYGENOMICS GENE DIAGNOSIS TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the amount of isolated DNA is too small, so it is only suitable for the detection of mutation sites, and it is not suitable for detecting changes in chromosome copy number.

Method used

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  • Kit, device and method for detecting copy number of embryo chromosomes
  • Kit, device and method for detecting copy number of embryo chromosomes
  • Kit, device and method for detecting copy number of embryo chromosomes

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0056] Example 1: A method for detecting the copy number of embryonic chromosomes

[0057] Step 1: Take maternal blood and prepare plasma

[0058] In this embodiment, a total of 5 maternal blood samples were taken, and the blood samples were centrifuged at high speed to obtain plasma samples from which blood cells were removed. The plasma volume of each sample was about 1ml, and the sample codes were: G367, G374, G379, G435 and G440. The above samples were all collected by Professor Wu Lingqian of Xiangya School of Medicine, Central South University.

[0059] Step 2: Extract plasma DNA

[0060] The DNA in the plasma can be extracted using the DNA extraction kit produced by Qiagen (Product No. 57704).

[0061] Step 3: Prepare plasma DNA for paired-end sequencing library

[0062] Fill in the end of the extracted DNA and phosphorylate the 5' end: mix 30 μl of DNA, 45 μl of pure water, 10 μl of T4 DNA ligase buffer with 10 mM ATP, 4 μl of 10 mM dNTP Mix, 5 μl of T4 DNA polymer...

Embodiment 2

[0086] Example 2: Another method for detecting embryo chromosomes

[0087] Another method for detecting embryonic chromosomes of the present invention is different from Example 1 in that we do not sequence all the sequences in the operation, but by physical methods, preferably by agarose gel electrophoresis Isolate the DNA at any point or any interval between 100bp-250bp, and sequence the DNA at any point or any interval between 100bp-250bp. The rest of the operations or calculation steps can be the same as those in Embodiment 1.

Embodiment 3

[0088] Example 3: A kit for detecting the copy number of embryonic chromosomes

[0089] Corresponding to the detection method in Example 1, the inventors of the present application have developed a kit that can be used to detect the copy number of embryonic chromosomes, which includes:

[0090] The equipment used to draw blood from the mother can be any blood collection needle, syringe, etc. that can be used to draw blood;

[0091] a device suitable for separating blood cells and plasma in said blood, which may be a microtube suitable for holding blood in a centrifuge or any other container or device suitable for separation;

[0092] The reagents and instruments for extracting DNA in the plasma may include: protease, saturated phenol, chloroform:isoamyl alcohol (24:1), sodium acetate, absolute ethanol, 70% ethanol, TE solution, etc. You can also choose the DNA extraction kit produced by Qiagen to extract the DNA in the plasma (the product number is 57704) and any other reagen...

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PUM

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Abstract

The invention relates to a kit, device and method for detecting the copy number of embryo chromosomes. By using the kit, device and method provided by the invention, the copy number of embryo chromosomes can be detected non-invasively and economically. The inventor finds that most embryo DNAs (deoxyribonucleic acids) in maternal plasma are 100bp-250bp segments and the ratio of each chromosome to the whole DNA is equal to the ratio of each chromosome to the DNA at any point or in any interval within the range of 100bp to 250bp in the maternal plasma. According to the method provided by the invention, the copy number of chromosomes to be detected can be determined by simply confirming which chromosome each DNA segment of the DNA at any point or in any interval within the range of 100bp to 250bp comes from, calculating the ratio of the number of DNA segments from the chromosomes to be detected to the number of DNA segments from reference chromosomes in all the DNAs at any point or in any interval within the range of 100bp to 250bp in the same sample, calculating the differences among the samples and determining according to the difference values.

Description

technical field [0001] The invention relates to a kit, device and method for detecting embryo chromosome copy number. Background technique [0002] Chromosomal copy number abnormalities are closely related to human diseases. On average, 9 out of every 1000 newborns will carry diseases caused by abnormal chromosome copy number (1) . Therefore, it is very important to be able to detect the chromosome copy number before the fetus is born. However, currently used diagnostic methods, including amniocentesis and chorionic villus sampling, are invasive methods that pose certain risks to pregnant women and fetuses. Using serum protein markers and ultrasound to detect whether the fetus has chromosomal copy number abnormalities is not invasive, but it does not directly detect the causative factors, so the accuracy and sensitivity are not very good (2) , there is also the problem of not being able to detect abnormal chromosome copy number diseases as early as possible. This situat...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68
Inventor 王珺张建光田凤高扬周代星
Owner HANGZHOU BERRYGENOMICS GENE DIAGNOSIS TECH
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