Method for detecting fetal thalassemia pathogenic gene and kit

Abstract

The invention discloses a method for detecting a fetal thalassemia pathogenic gene, which comprises the following steps of: (1) screening SNP sites, wherein the SNP sites are used for designing a primer pool of the thalassemia gene in an amplification genome and for capturing the probe of the thalassemia gene of the free DNA in the plasma of a pregnant woman; (2) extracting the free DNA in the plasma of the pregnant woman and the whole blood genomic DNA of the father, the mother and a born sibling and constructing a corresponding DNA library, and carrying out template preparation and enrichment; (3) sequencing the free DNA and the whole blood genomic DNA library in step (2); (4) constructing the haploid genotype of the SNP sites on the thalassemia gene, combining the sequencing informationof the free DNA and the whole blood genomic DNA library, analyzing the genetic condition of the parent source and the genetic condition of the parent source, so as to determine the corresponding genotype of the SNP sites of the fetal. According to the method, target area capture and high-throughput sequencing technology are used, so that the noninvasive antepartum detection of the thalassemia isrealized; the required sample amount is small; on the basis of detecting the mutation of the parent source, the method can realize the detection of the gene mutation of the maternal source of the fetal.

Description

technical field [0001] The invention relates to the technical field of gene detection, in particular to a method and a kit for detecting fetal thalassemia-causing genes. Background technique [0002] Thalassemia (referred to as thalassemia) is one of the high-incidence genetic diseases in the world. It is due to the mutation or deletion of human α, β-globin gene, which leads to the imbalance of α, β-globin peptide chain synthesis rate, which causes hemolysis. sexual anemia. The two common types of thalassemia are α-thalassemia and β-thalassemia, α-thalassemia-related genes are HBA2 and HBA1, and β-thalassemia-related genes are HBB. Thalassemia is concentrated in tropical and subtropical regions, mostly in Mediterranean countries, followed by the Middle East, India, Pakistan, Southeast Asia, southern China and North Africa. The United States is an immigrant country, and its incidence is also relatively high. The provinces south of the Yangtze River in my country are high-in...

AI Technical Summary

Problems solved by technology

However, these methods are based on single-site PCR technology. Due to the low content of plasma DNA and the characteristics of high fragmentation, it is likely to cause false negatives in PCR.

In addition, these methods can only detect the presence of specific paternal mutations that are different from the mother, but cannot further confirm the presence of maternal mutations

Images