Thalassemia gene detection method based on high-throughput sequencing technology

Abstract

The invention discloses a thalassemia gene detection method based on a high-throughput sequencing technology. The thalassemia gene detection method mainly comprises the following steps that alpha & beta-thalassemia related gene fragments are specifically amplified based on PCR amplification primers of a span breakpoint and a mutation site designed by adopting a Gap-PCR method, library establishment is conducted on PCR products, library products are subjected to high-throughput sequencing, sequencing data uses a human genome hg19 as a reference sequence, sequencing depth values of gene loci in a target area chr16: 215000-236000 are analyzed according to a sequence alignment score Q10, and alpha-thalassemia deletion types are determined according to the distribution of the sequencing depth values of different loci; meanwhile sequence alignment is performed through the target area chr16: 215000-236000 and a target area chr16: 5246400-5248600, and alpha & beta-thalassemia mutation types are determined according to the basic types of specific sites. By the adoption of the thalassemia gene detection method, simultaneous detection of 6 mutation genetypes of alpha-thalassemia and 26 mutation genetypes of beta-thalassemia can be achieved.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a method for detecting thalassemia genes based on high-throughput sequencing technology. Background technique [0002] Thalassemia (Thalassemia), referred to as thalassemia. Thalassemia is one of the most common monogenic genetic diseases, which are divided into α-thalassemia and β-thalassemia, which are caused by the loss or mutation of α or β globin gene, which leads to the disorder of globin chain synthesis, resulting in α chain and β chain The ratio is out of balance, and the relatively excess chains are in a free state and are prone to oxidative denaturation. The denatured chains are deposited in the red blood cells and attached to the intima, which reduces the deformability of the red blood cells, changes the membrane permeability and is easily destroyed, leading to hemolytic anemia. The α-thalassemia-related genes are HBA2 and HBA1, and the β-thalassemia-related genes are HB...

AI Technical Summary

Problems solved by technology

At present, there is no cure for thalassemia, and it can only be treated by traditional methods, that is, blood transfusion is the main treatment, which is expensive. Therefore, screening for thalassemia carriers is of great significance

[0003] At present, the main method for detecting α-deficient thalassemia is the Gap-PCR method, and the main method for detecting α-mutant thalassemia and β-mutant thalassemia is the PCR-RDB method, so the deletion of α&β-thalassemia cannot be detected Simultaneous detection with mutant type

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