Method for detecting alpha-thalassemia and beta-thalassemia point mutation

Abstract

The invention discloses a method for detecting alpha-thalassemia and beta-thalassemia point mutation, and belongs to the technical field of gene detection. The method provides a probe pair composed of different-tail double-stranded fluorescent probes, the different-tail double-stranded fluorescent probes labeled by different fluorescent groups are added into a PCR reaction system containing a to-be-detected sample according to a probe coding mode, asymmetric PCR amplification is utilized to generate a large number of single-stranded DNA templates, fluorescent chains are specifically combined with the single-stranded DNA templates to release fluorescence signals, the fluorescence net increment of different fluorescence channels is calculated, comparing the fluorescence net increment with a mutation judgment threshold value, and a point mutation type is judged. According to the method, asymmetric PCR and fluorescent chain detection are combined, the operation is easy and convenient, the detection time is short, an automatic detection process is compatible, the method is suitable for detection of a large number of samples, especially 24 thalassemia point mutation is detected through four fluorescence channels in a fluorescence coding mode, detection flux and mutation sites can be improved, and the use amount of reaction reagents and the use amount of samples are reduced.

Description

technical field [0001] The invention relates to a method for detecting alpha-thalassemia and beta-thalassemia point mutations, belonging to the technical field of gene detection. Background technique [0002] Thalassemia, also known as thalassemia, is more common in the Mediterranean, Southeast Asia and other regions. It is a fatal and disabled hereditary blood disease that seriously threatens human health. Since the early reported cases were almost all immigrants from the Mediterranean region, the disease was named thalassemia, or thalassemia for short. According to the classification of gene defects, it is mainly divided into α-streptoglobin thalassemia and β-streptoglobin thalassemia clinically. According to the genotype, clinically divided into α-thalassemia, β-thalassemia, γ-thalassemia and γβ-thalassemia, etc., the clinically high incidence is mainly the former two. α static type and standard type, and β thalassemia heterozygotes are generally called α or β thalassem...

AI Technical Summary

Problems solved by technology

Gene chip technology is to arrange a large number of oligonucleotide fragments or DNA fragments in an orderly manner on a solid-phase carrier. After the DNA of the sample to be tested is hybridized with the chip, the fluorescence intensity of the chip is scanned to obtain relevant information about the gene sequence and expression level in the sample. The degree of standardization and automation is high, and the quality control is convenient, but the detection price is high, and the data analysis and processing are complicated

High-throughput sequencing technology, also known as next-generation sequencing (NGS), can comprehensively analyze target DNA in one experiment, including single-base variation, short insertion-deletion, copy number variation, and structural variation. Disease genetic testing is widely used, but the testing price is high, the testing process is complicated, and data analysis requires specialized personnel and technical platforms

In summary, the existing thalassemia gene detection technology still cannot meet the requirements of economical, simple and rapid clinical diagnosis

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