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Amplification composition for detecting abnormal number of chromosomal aneuploid and rapid detection kit

An aneuploidy and chromosome technology, applied in the field of biotechnology detection, can solve the problems of small number, insufficient detection efficiency and accuracy, and small number of loci

Active Publication Date: 2015-05-27
BEIJING MICROREAD GENE TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The above-mentioned patents often use unreasonable or small number of STR sites, and the number of sites contained in each QF-PCR amplification system is small, resulting in insufficient detection efficiency and accuracy

Method used

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  • Amplification composition for detecting abnormal number of chromosomal aneuploid and rapid detection kit
  • Amplification composition for detecting abnormal number of chromosomal aneuploid and rapid detection kit
  • Amplification composition for detecting abnormal number of chromosomal aneuploid and rapid detection kit

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Embodiment 1

[0212] The following are the DNA samples of 1 patient with trisomy 21, 1 patient with trisomy 18, 1 patient with trisomy 13, 1 patient with Klinefelter syndrome and 2 normal persons using the present invention specific implementation.

[0213] 1. DNA extraction

[0214] Chelex-100 and DNA extraction kit (Tiangen Biochemical) were used for DNA extraction. The operation steps were in accordance with the instructions. After the DNA extraction was completed, the DNA was quantified with a UV spectrophotometer and diluted to 2 ng / μl.

[0215] 2.PCR

[0216] 2.1 Reaction system:

[0217] A total of 27 pairs of primers for 23 21, 18, 13, X chromosome STR loci and 4Y chromosome loci were dissolved respectively and made into a working solution with a concentration of 10 μM, and then made into a primer mixture (Primer mix ): See the sequence list in the appendix for the primer sequence.

[0218] Table 2 The volume ratio of each primer

[0219] site name

Volume (μl)

...

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Abstract

The invention relates to an amplification composition for detecting abnormal number of chromosomal aneuploid and a rapid detection kit, and belongs to the technical field of biology. The amplification composition can be used for amplification detection of STR sites related to 8 21# chromosomes, 6 18# chromosomes, 6 13# chromosomes and 3 X chromosomes and amplification detection of four amelo; all sites can be amplified by a quantitative fluorescent PCR (QF-PCR) technology, so as to detect the abnormal number of 21, 18, 13, X and Y chromosomal aneuploid. According to the kit, detection cycle is short, so the result can be obtained within 5 hours after obtaining the sample; the sensitivity is high; DNA at nanogram level, which is equal to DNA contained in 100 cells, is needed for each detection, so few samples are needed; 1 to 2ml of amniotic fluid can be collected to meet the demand; due to high sensitivity, a chimera containing more than 10% of trisomies can be detected.

Description

technical field [0001] The invention relates to the field of biotechnology detection, and is a detection method based on quantitative fluorescent PCR (Quantitive Fluoresent PCR, QF-PCR) at the DNA level, which can detect common trisomy syndromes 21, 18, 13 and X, Y sex chromosomes Fast, easy and accurate detection of abnormal numbers. Background technique [0002] Chromosome abnormalities and structural aberrations are one of the important causes of birth defects (Gardner R J M, Sutherland G R. Chromosome abnormalities and genetic counseling [M]. New York: Oxford University Press, 1996: 300-320.), chromosome incomplete Ploidy is an abnormal number of chromosomes, including haplotype, trisomy, polysomy and mosaicism. Clinically common include trisomy 21 (Down's syndrome), trisomy 18 (Edward syndrome), trisomy 13 (Patau syndrome), and some sex chromosome aberrations, such as Klinefelter syndrome (47,XXY) and Turner syndrome (45, X) and so on. Studies have shown that 13, 18,...

Claims

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Application Information

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IPC IPC(8): C12Q1/68C12N15/11
CPCC12Q1/6827C12Q1/6883C12Q2600/156
Inventor 张晔宋欣欣张晓娜吕悦心陈初光
Owner BEIJING MICROREAD GENE TECH
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