Method for detecting fetal patrilineal DNA point mutation from pregnancy plasma

A point mutation, plasma technology, applied in the field of DNA detection, can solve the problems of expensive instruments, limited application, affecting diagnostic results, etc., and achieves the effect of reliable method and high sensitivity

Inactive Publication Date: 2007-06-06
THE THIRD AFFILIATED HOSPITAL OF THIRD MILITARY MEDICAL UNIV OF PLA
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  • Abstract
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Problems solved by technology

However, this method has the following shortcomings: as we all know, the influence of detection reliability is determined by the type of point mutation, the content of target DNA in the sample, and the ratio between target DNA and background DNA.
The technology platform is based on single base extension reaction (SBER) with high specificity and matrix assisted laser desorption ionization/time-of-flight mass spectrometry (matrix assisted laser desorption ionization-time offlight MS, MALDI-TOF- MS), will have the potential to be applied to non-invasive prenatal diagnosis, but this m

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Embodiment Construction

[0022] The present invention will be further described below in conjunction with embodiment:

[0023] The steps of the present invention are as follows:

[0024] (1), sample processing: extract the peripheral blood of pregnant women, centrifuge to separate the plasma, and extract DNA from the plasma;

[0025] (2), gap-ligase chain reaction (Gap-LCR): the DNA in step (1) is used as a template to carry out gap-ligase chain reaction (Gap-LCR), wherein, as the four Gap-LCR primers Among oligonucleotide fragments p1, p2, p3, p4, primers p1, p3 bind to one DNA strand, primers p2, p4 bind to another DNA strand, and primers p1, p3 have the following characteristics:

[0026] Primer p1: Primer p1 is an upstream primer, and its 5′ end is provided with a DNA sequence with low homology to human DNA as a recognition sequence for fluorescent quantitative PCR;

[0027]Primer p3: Primer p3 is a downstream primer, and its 3′ end is provided with a DNA sequence that has low homology to human ...

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Abstract

A method to detect the father DNA dot mutation of the fetus from the gravida plasma includes three processes: the sample treatment, recognizing the dot mutation by the space-connecting enzyme reaction, detecting the connecting product by the fluorescent quantitative PCR. The method is reliable and has the high sensitivity, so it can detect the father DNA dot mutation of the fetus from the gravida plasma accurately.

Description

(1) Technical field [0001] The invention relates to a DNA detection method, in particular to a method for detecting free DNA point mutations. (2) Technical background [0002] With the transformation of my country's population and disease patterns, the problem of birth defects has become increasingly prominent, and has become the main cause of newborn and infant deaths, and disability of children and adults in my country. According to the statistics of the Ministry of Health, the incidence of birth defects is about 13.05‰, of which the proportion of birth defects caused by genetic factors is as high as 25%. There is no effective treatment for hereditary birth defects. At present, prenatal diagnosis can be used to effectively avoid the birth of children, which is conducive to prenatal and postnatal care and improving the quality of the population. [0003] The traditional amniocentesis and chorionic villus extraction are the prenatal diagnostic techniques commonly used in ma...

Claims

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Application Information

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IPC IPC(8): G01N21/76G01N21/64C12Q1/68
Inventor 易萍李力陈竹钦郭建新
Owner THE THIRD AFFILIATED HOSPITAL OF THIRD MILITARY MEDICAL UNIV OF PLA
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