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Reagent box for detecting No 21 chromosome and idiochromosome number abnormality

A chromosome number and chromosome technology, which is applied in the field of kits for detecting abnormal number of chromosomes 21 and sex chromosomes in clinical samples, can solve the problem that there is no effective prenatal screening method for sex chromosome abnormalities.

Active Publication Date: 2008-12-17
DAAN GENE CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

There is currently no effective prenatal screening method for targeted chromosomal abnormalities

Method used

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  • Reagent box for detecting No 21 chromosome and idiochromosome number abnormality
  • Reagent box for detecting No 21 chromosome and idiochromosome number abnormality
  • Reagent box for detecting No 21 chromosome and idiochromosome number abnormality

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0046] Embodiment 1: detection kit and its use

[0047] (1) Prepare a kit including the following components: 1 tube of PCR reaction solution (800 μl / tube), 1 tube of primer mix (200 μl / tube), 1 tube of Taq enzyme (25 μl / tube), 1 tube of positive trisomy 21 Tube (25ul / tube), 1 tube (25μl / tube) of sex chromosome abnormality positive standard, 1 tube (25μl / tube) of negative control.

[0048] (2) Specimen collection, transportation and storage:

[0049] 1. Specimen collection: The specimens are blood, amniotic fluid, and villi tissue. The blood is 2ml of venous blood or 0.5-1ml of fetal umbilical cord blood, which is anticoagulated with EDTA; 2-5ml of amniotic fluid or two pieces of villous tissue are obtained by puncture.

[0050] 2. Storage: It can be detected immediately, stored at 4°C for one week, and stored at -20°C for one year.

[0051] 3. Transportation: Specimens should be transported using 0°C curling.

[0052] (3) Detection steps and result analysis:

[0053] For...

Embodiment 2

[0062] Example 2: Detection of trisomy 21 by QF-PCR amplification of the STR locus on chromosome 21

[0063] Blood samples, amniotic fluid or villi tissues from donors were subjected to DNA extraction and purification according to the standard procedure of Qiagen DNA extraction kit. The concentration of each sample DNA solution was adjusted to 20-40ng / μl with TE buffer (5mM Tris-HCl pH8.0, 1mM EDTA pH8.0). Use the primer mixture provided in the kit, PCR buffer system and Taq enzyme to carry out the amplification reaction according to the detection protocol of the kit and load the sample for analysis. The amplification reaction simultaneously amplifies seven sites including D21S11, D21S1435, D21S1411, AMXY, DXS981, DXS6809, and X22. The analysis pattern of the loading results is as follows: image 3 , Figure 4 . As shown in the figure, the genotypes of the three loci D21S11, D21S1435 and D21S1411 on chromosome 21 are amplified to analyze the genotype of the loci, and when i...

Embodiment 3

[0064] Example 3: Detection of Klinefelter Syndrome by QF-PCR Amplification of Genetic Loci on Sex Chromosomes

[0065] Blood samples, amniotic fluid or villi tissues from donors were subjected to DNA extraction and purification according to the standard procedure of Qiagen DNA extraction kit. The concentration of each sample DNA solution was adjusted to 20-40ng / μl with TE buffer (5mM Tris-HCl pH8.0, 1mM EDTApH8.0). Use the primer mixture provided in the kit, PCR buffer system and Taq enzyme to carry out the amplification reaction according to the detection protocol of the kit and load the sample for analysis. The amplification reaction simultaneously amplifies seven sites including D21S11, D21S1435, D21S1411, AMXY, DXS981, DXS6809, and X22. The analysis pattern of the loading results is as follows: Figure 5 . As shown in the figure, three loci D21S11, D21S1435 and D21S1411 on chromosome 21 amplify and analyze the genotypes of the loci, forming a 1:1 fluorescence peak (area...

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Abstract

The invention relates to a kit used for screening the numerical abnormality of 21-trisomy and sex chromosome at early stage. The kit adopts quantitative fluorescence multiplex polymerase chain reaction technology, carries out the fluorescent primer sevenfold multiplex amplification respectively to differential genetic locus on the 21-trisomy and sex chromosome and analyzes the numerical abnormality of chromosome according to the difference of the dosage of amplified products, thus achieving the purpose of detecting the numerical abnormality of 21-trisomy and sex chromosome.

Description

[0001] Field [0002] The invention relates to a kit for detecting abnormal numbers of chromosome 21 and sex chromosomes in clinical samples, in particular to a kit for early screening of trisomy 21 and abnormal numbers of sex chromosomes by quantitative fluorescent multiple polymerase chain reaction technology. Background of the invention [0003] Trisomy 21 is also known as Down syndrome (DS). The incidence of Down syndrome in newborns is about 1 / 800. It is estimated that more than 600,000 children with DS are born in my country at present, that is, one child with DS is born every 20 minutes on average. The incidence of DS increases with the increase of mother's childbearing age, especially when the mother's age is greater than 35 years old, the incidence rate increases significantly. Clinically, for women of advanced age (women over 35 years old) in the childbearing period, the ovaries are affected by various harmful substances and radiation, which increases the chance of...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68
Inventor 陈华云李明江帆章均李佩琼
Owner DAAN GENE CO LTD
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