Method of simultaneously detecting the numbers of human chromosomes 21 and 18

A chromosome number and chromosome technology, applied in the field of molecular biology, can solve the problems of different results, wrong judgment, affecting the correctness of the results, etc., to achieve objective and accurate results and reduce interference.

Inactive Publication Date: 2011-06-08
广州医学院第三附属医院
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However, some factors in the management experiment, such as the difference between wells, the difference in template loading, etc., can lead to completely different results, resulting in wrong judgments
In addition, Lu Shiming et al. disclosed the method of using real-time quantitative PCR technology to detect the number of human chromosome 21 in the patent application with the application number 200510049601.2 and the title of invention "method for detecting the

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  • Method of simultaneously detecting the numbers of human chromosomes 21 and 18

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[0023] A kind of multiple real-time fluorescent PCR relative quantitative method described in the present invention simultaneously detects the method for the number of human chromosome 21 and 18 in the same reaction tube, comprising the following steps:

[0024] (1) DNA extraction:

[0025] The DNA of the sample was extracted according to the conventional phenol-chloroform method, for example, using the QIAamp DNA BloodMini Kit kit (Qiagen, Germany), extracting the DNA according to the instructions, and detecting the DNA concentration and purity with a micro-nucleic acid protein detector (NanoDrop ND-1000). Required DNA purity requires OD 260 / OD 280 The value of 1.8 ~ 2.0;

[0026] (2) Design and synthesis of primers and probes:

[0027] The primer sequences designed to select a partial fragment of the amyloid precursor protein gene (APP) on chromosome 21 21q21.3 are:

[0028] Upstream primer: 5′-CCCAGGAAGGAAGTCTGTACCC-3′SEQ ID NO.1;

[0029] Downstream primer: 5′-CCCTTG...

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Abstract

The invention relates to a method of simultaneous detecting the numbers of human chromosomes 21 and 18 utilizing multiple and real time relative quantitative fluorescence PCR. A relative quantitative method using real time fluorescence PCR technology to compare CT values is applied, which is deta CT value method. Partial fragments of APP and TYMS genes on chromosomes 21 and 18 are internal mutual reference in the same tube, and are amplified simultaneously; deta CT value of the two amplification products is calculated; the relative value of deta CT to template is adjusted; and the deta CT value is used to determine the numbers of human chromosomes 21 and 18. The invention possesses the advantages of: automatic closed tube operation, simplicity, rapid speed, low cost, no pollution, large flux, high specific and sensitive detection, reliable accuracy, and simultaneous detection of the number of human chromosomes 21 and 18.

Description

technical field [0001] The invention belongs to the field of molecular biology, and relates to a method for detecting the number of chromosomes, in particular to a method for simultaneously detecting the number of human chromosomes 21 and 18 in the same reaction tube by using a multiple real-time fluorescent PCR relative quantitative method. Background technique [0002] Trisomy 21 and 18 are the two most common chromosomal diseases of fetal birth defects. The incidence rates are 1 / 600-1 / 800 and 1 / 6000 respectively. On the basis of the two, there is one more (or part of the 21 / 18 translocation chromosome). Patients with these two diseases have severe mental retardation and multiple deformities, and the early mortality of children is high, which brings heavy burdens to families and society. There is no effective treatment for these two diseases, and prenatal diagnosis is mainly used to avoid the birth of defective children. At present, it mainly relies on the traditional cy...

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Application Information

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IPC IPC(8): C12Q1/68G01N21/64
Inventor 孙筱放眭建忠
Owner 广州医学院第三附属医院
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