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Chromosome specific loci screening method and application thereof

A technology of specific sites and screening methods, applied in biochemical equipment and methods, microbial measurement/inspection, etc., can solve the problems of large sequencing volume, high price, and unfavorable promotion, and achieve low cost, small quantity, and improved detection The effect of the flux

Active Publication Date: 2013-07-24
SUZHOU BASECARE MEDICAL DEVICE CO LTD
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  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, this technology still has certain deficiencies. Since this technology needs to detect the entire genome DNA fragment, the gene sequence of the entire human genome has more than 3 billion bases. To sequence all these bases requires a large amount of sequencing. It is relatively large, resulting in a relatively high price for prenatal testing, which is not conducive to large-scale promotion. For example, the current non-invasive DNA prenatal testing using Illumina's Hiseq2000 sequencer can complete up to 12 samples in one lane, and the testing price of one sample is 5,000 diverse

Method used

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  • Chromosome specific loci screening method and application thereof
  • Chromosome specific loci screening method and application thereof

Examples

Experimental program
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Embodiment 1

[0052] A screening method for specific chromosome sites, comprising the following steps:

[0053] Step 1. Preliminary screening of chromosome-specific sites: According to the human genome sequence (GRCH Build37) of the National Center for Biotechnology Information (NCBI), a preliminary screening of a chromosome-specific site to be studied will yield 200,000 to 300,000 screening sites point sequence, the primary screening point sequence meets the following conditions:

[0054] (1) The value range of the sequence length is 50-70bp;

[0055] (2) GC content is between 45% and 55%;

[0056] (3) The character "N" is not included in the sequence;

[0057] (4) Compared with the single nucleotide polymorphism site database (dbSNP) of the National Center for Biotechnology Information (NCBI), there is no single nucleotide polymorphism site (SNP) in the sequence;

[0058] (5) Compared with the copy number variation (CNV) information in the genome variation database (DGV) of the Nationa...

Embodiment 2

[0079] For the application of the screening method of the above-mentioned chromosomal specific loci in prenatal detection, see figure 1 , including the following steps:

[0080] (1) Sample preparation

[0081] 5-10ml of peripheral blood was drawn from 200 pregnant women over 12 weeks pregnant, of which 5 were pregnant with trisomy 21, 2 were pregnant with trisomy 18, and 1 was pregnant with trisomy 13. The plasma was collected by centrifugation. Separation, using QiAamp Blood Mini Kit kit to extract free DNA in plasma;

[0082] (2) Separation of site DNA fragments

[0083] According to the 2000-3000 chromosome-specific sites obtained in Example 1, randomly select 500-1000 specific site sequences from chromosomes 21, 18, and 13 respectively, and design probes, combined with biomagnetic beads, to capture the target DNA The following steps are taken specifically: designing probes according to the specific site sequence; binding the probes to biomagnetic beads; adding the free ...

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Abstract

The invention provides a chromosome specific loci screening method comprising the steps that: chromosome specific loci preliminary screening is carried out; chromosome specific loci comparison and removing are carried out; chromosome specific loci screening is carried out with a PCR primer design method; the loci screened by the PCR primers are finely screened with a real-time quantitative PCR reaction; and the like. The invention also provides the application of the screening method in prenatal testing. According to the chromosome specific loci screening method provided by the invention, specific locus sequence is screened on a specific chromosome by using common computer software. The operation is simple, cost is low, and the specific loci obtained by screening have small number and high specificity.

Description

technical field [0001] The invention belongs to the field of biotechnology, relates to a screening method for specific chromosome sites, and also relates to the application of the screening method, such as prenatal detection. Background technique [0002] my country is a country with a large population and a high incidence of birth defects. According to the latest "Report on the Prevention and Control of Birth Defects in my country (2012)" released by the Ministry of Health, birth defects have increasingly become a prominent public health and social problem in China. At present, about 16 million newborns are born in my country every year, and the incidence rate of birth defects is about 5.6%, that is, about 900,000 new cases of birth defects are added every year. [0003] The main cause of a large number of birth defects is chromosomal abnormality, which mainly refers to abnormalities in the number or structure of chromosomes. Quantitative abnormalities refer to changes in...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68
Inventor 梁波孔令印
Owner SUZHOU BASECARE MEDICAL DEVICE CO LTD
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