Thalassemia mutant gene detection primer pair and kit, and application and library construction methods thereof

Abstract

Disclosed are a thalassemia mutant gene detection primer pair and kit, and application and library construction methods thereof. The thalassemia mutant gene detection primer pair comprises primer sequences shown as SEQ ID NO: 1-8. The thalassemia mutant gene detection primer pair can be applied to simultaneously detecting a number of mutation types and help discover new mutation types, and when combined with second-generation sequencing, can greatly reduce the detecting cost and improve throughput, thereby achieving significant cost advantages and being beneficial to popularized application ofthalassemia mutant gene detection.

Description

Technical field [0001] The present invention relates to the technical field of thalassemia detection, in particular to a primer set, a kit, its application and a library construction method for thalassemia mutant gene detection. Background technique [0002] Thalassemia (hereinafter referred to as thalassaemia) is a common hemolytic single-gene genetic disease that occurs mostly in the Middle East, Central Asia, Africa, Southeast Asia, and South China. The molecular mechanism of thalassaemia is that the defect of the globin gene reduces or lacks the synthesis of one or more of the peptide chains that it encodes, resulting in an imbalance in the proportion of hemoglobin components, which in turn leads to hemoglobin instability. According to different types of defective globin genes, thalassaemia is mainly divided into α-thalassaemia and β-thalassaemia. [0003] Alpha-thalassemia is a hemolytic anemia disease caused by the deletion or mutation of the alpha-globin gene that inhibits ...

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Problems solved by technology

[0009] All of the above-mentioned thalassemia gene detection methods have shortcomings to varying degrees. For example, the Gap-PCR method can only detect deletion types; among various detection methods for mutation types, PCR-ARMS can detect few types of mutations, and PCR-ASO, Although PCR-RDB and real-time fluorescent quantitative PCR methods are low in detection cost and can quickly detect multiple mutation types, they can only detect common hotspot mutations (for example, detect 15-20 mutation types in China), and cannot detect other rare mutation types and New mutations, there is a certain rate of missed detection

The PCR reaction combined with sequencing method can not only detect multiple mutation types, but also discover new mutation types, which is the best choice in terms of detection results, but it also has the disadvantage of high cost.

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