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Down's syndrome gene diagnostic new technology and its use

A technology for Down syndrome and genetic diagnosis, applied in the field of genetic diagnosis of diseases, can solve problems that have not yet been seen

Inactive Publication Date: 2005-10-19
刘敬忠
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

There is no report on D.S. molecular diagnosis using SYBR-Q-PCR combined with D.C. analysis technology

Method used

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  • Down's syndrome gene diagnostic new technology and its use
  • Down's syndrome gene diagnostic new technology and its use
  • Down's syndrome gene diagnostic new technology and its use

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0025] Detection of Chromosome Number 21 by Double Tube Relative Quantitative Method

[0026] 1. Albumin gene as a reference gene, primer sequence, Alb1: 5′-GCT GTC ATC TCT TGT GGGCTG T-3′;

[0027] Alb2: 5′-ACT CAT GGG AGC TGC TGG TTC-3′;

[0028] 2. Select the DSCR3 gene on chromosome 21 as the target gene, primer sequence, 21 # -1:5′-GTGGAG GGA AGT CAT GCA TTT-3; 21 # -2: 5'-AAC ATT GAC TGT GGC TCTTGC-3. '

[0029] 3. The SYBY-Q-PCR reaction mixture contains: 2.5μl 10×SYBY PCR Buffer; 1.5mMMgCl2; 200mMdNTPs; 300nM each of primer Alb1, Alb2 or 21 # -1, 21 # -2; 0.6U TaqDNA polymerase, 50ng of DNA. The total volume is 50 μl / PCR.

[0030] 4. The thermal cycle is performed on a quantitative PCR instrument such as GeneAmp 5700 or ABI 7000. 95°C, 10min→(95°C, 15s---60°C, 1min), 20-30 cycles, and then conduct a 60°C-95°C melting curve analysis.

[0031] 5. Results

[0032] The fluorescence amplification curves and D.C obtained by detecting the DNA of normal people with t...

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Abstract

The present invention discloses new gene diagnosis and antepartum diagnosis technology of Down's syndrome. The present invention performs real-time PCR with fluorescent dye combined with deliquescence curve analysis, and the relative quantitative analysis on the reference gene and the target gene on the 21st. chromosome to determine the normality of 21st. chromosome to be detected. The technology may be used in the gene diagnosis of patient. The present invention has high detection specificity, high accuracy and reliability, high sensitivity, no need of post-treating PCR product and other advantages.

Description

technical field [0001] The present invention relates to a gene diagnosis method and application of a disease, in particular to a new technique for gene diagnosis of Down's syndrome and its application. Background technique [0002] Abnormal number of human chromosomes leads to congenital diseases. The most common is Down’s Syndrome (D.S.), with an incidence of about 1 / 800. It is caused by an extra chromosome 21, also known as Trisomy 21. Children with mental retardation, low, have special faces and so on. In addition, there is 18-trisomy, and 13-trisomy has no effective treatment so far. Pregnant women, especially elderly pregnant women, should be screened during pregnancy. For high-risk pregnant women screened out, amniotic fluid should be taken in the second trimester for amniotic fluid cell culture and chromosome examination to make a diagnosis. However, the karyotype analysis technology is time-consuming and laborious, the results are slow, and the operator's technical...

Claims

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Application Information

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IPC IPC(8): C12Q1/68
Inventor 刘敬忠
Owner 刘敬忠
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