Method of concentrating fetus dissociative DNA in pregnant woman blood plasma

A plasma, pregnant woman technology, applied in DNA preparation, recombinant DNA technology, DNA/RNA fragments, etc., can solve the problem of low fetal DNA

Inactive Publication Date: 2007-08-15
陈熹
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  • Claims
  • Application Information

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Problems solved by technology

However, the content of fetal DNA in maternal blood is low, and due to the influence of the strong DNA background of the mother, the current research on prenatal diagnosis based on fetal cell-free DNA is mostly limited to paternal genetic diseases

Method used

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Embodiment Construction

[0004] 30 cases of mid-pregnancy women from January 2005 to October 2005 were selected, and 6ml of peripheral blood was collected respectively. Wherein 3ml, use qiagen whole blood DNA extraction kit to extract DNA after the method of the present invention is processed. Get pregnant woman's peripheral blood 3-5ml, add EDTA and analyze pure formaldehyde, make its final concentration be respectively 0.5M and 1ml / L. Within 24 hours, centrifuge at 1600g for 10 minutes at room temperature. The plasma was transferred to a new centrifuge tube, and the above steps were repeated. The plasma was extracted with a qiagen extraction kit for DNA. The other 3ml was directly extracted with a qiagen extraction kit for DNA.

[0005] The fluorescent quantitative PCR method was used to amplify the SRY gene to obtain the relative concentration of fetal DNA in total DNA in the plasma of pregnant women. In the non-enriched DNA treatment group, the relative concentration was 6.10%. In the treatment ...

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Abstract

The invention discloses an agent to enrich franc free DNA in pregnant woman plasma and method, which comprises the following steps: making the agent include 0.5M EDTA(pH=8.0) and analytical pure methylene oxide; allocating 3-5 ml peripheral blood; adding into EDTA and analytical pure methylene oxide; making the final density at 0.5 M and 1ml / L; centrifuging 1600g under room temperature at 10 min in 24 h; transferring the plasma to a new centrifuge tube; repeating the steps; can being used to successive trials or -80 cold conservation. This invention can be used to miscellaneous brand DNA extraction, which can increase the extracting content of pregnant woman plasma apparently.

Description

technical field [0001] The invention relates to a reagent for enriching fetal free DNA in pregnant woman's plasma and a method for using the reagent to enrich fetal free DNA in pregnant woman's plasma. Background technique [0002] At present, prenatal diagnosis mainly uses fetal tissue to detect some genetic diseases, which has very important clinical significance for the prevention and control of neonatal morbidity and mortality. However, intrusive diagnostic methods such as amniotic fluid and chorionic villus puncture have the opportunity to cause certain harm to the fetus and pregnant woman. Therefore, these methods are usually reserved for high-risk pregnant women who are suspected of carrying a fetus with a genetic disorder. In view of the limitations of traumatic prenatal diagnosis, the method of non-traumatic prenatal diagnosis has great development space and potential. Among them, the phenomenon of cell interpenetration between mother and fetus has been confirmed ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C07H21/04C12N15/11C12N15/10
Inventor 陈熹
Owner 陈熹
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