A method for detecting fetal dna content in plasma of pregnant women

Abstract

The invention discloses a method for detecting the fetus DNA content of pregnant woman blood plasma. The method comprises the following steps: simultaneously carrying out multiple fluorescence PCR amplification and product capillary electrophoresis separation on multiple SNP sites in the genome and the plasma of a pregnant woman through multiple fluorescence PCR amplification and capillary electrophoresis separation based on insertion / deletion SNP sites which are different in mother genome and fetus genome; selecting SNPs homozygous in the maternal genome and heterozygous in the fetus, carrying out peak height correction on the obtained capillary electrophoresis detection result by using a reference sample, and correcting the detection background by using the maternal genome DNA in order to quantitatively analyze two allele fragments of many SNP sites in the blood plasma DNA and obtain the proportion of the fetus DNA in the mother blood plasma DNA. The method has the advantages of simplicity, fastness, low cost and high accuracy when used in high-flux detection of the multiple SNP sites, and has wide application prospect in the field of noninvasive antenatal diagnosis.

Description

technical field [0001] The invention belongs to the technical field of life sciences, and in particular relates to a method for detecting fetal DNA content in plasma of pregnant women. Background technique [0002] With the deepening understanding of the molecular basis of genetic diseases, the technical ability to detect specific gene mutations has been continuously improved, providing a theoretical and technical basis for the development of non-invasive prenatal testing based on DNA testing. In 1997, Lo et al. used polymerase chain reaction (PCR) to discover the presence of fetal DNA in the plasma of pregnant women for the first time. Soon other scholars also confirmed the existence of fetal free DNA (cffDNA) and pointed out its potential clinical application value. Cell-free fetal DNA is thought to originate from lysis of placental trophoblast cells or release from naturally apoptotic fetal cells. Lo's team found that 59% of pregnant women could detect fetal cell-free DN...

AI Technical Summary

Problems solved by technology

[0010] In order to overcome the limitation that the existing detection technology based on epigenetic characteristics requires epigenetic specific sites and the defects of high-throughput sequencing detection technology that is time-consuming, complicated to operate and expensive, the present invention aims to provide a pregnant woman The detection method of fetal DNA content in plasma can accurately calculate the proportion of fetal DNA when the mother's genome DNA is available; when the mother's genome DNA is not available, it can also estimate the proportion of fetal DNA

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