Multi-PCR primer for detecting beta-mediterranean anemia mutation based on next-generation sequencing technology and method and application

Abstract

The invention provides a multi-PCR primer for detecting beta-mediterranean anemia mutation based on the next-generation sequencing technology, a method and application. The multi-PCR primer includes two or more primer pairs, namely, the primer pairs shown in SEQ ID NO:1 and SEQ ID NO:2, the primer pair shown in SEQ ID NO:3 and SEQ ID NO:4, the primer pair shown in SEQ ID NO:5 and SEQ ID NO:6, the primer pair shown in SEQ ID NO:7 and SEQ ID NO:8, the primer pair shown in SEQ ID NO:9 and SEQ ID NO:10 and the primer pair shown in SEQ ID NO:11 and SEQ ID NO:12. The multi-PCR primer can be used for efficiently detecting beta-mediterranean anemia mutation diversity and can cover 46 point mutation of beta-mediterranean anemia mutation of people in south of China.

Description

technical field [0001] The invention belongs to the field of molecular biology, and in particular relates to a multiplex PCR primer, method and application for detecting beta-thalassemia mutations based on next-generation sequencing technology. Background technique [0002] β-thalassemia is a monogenic genetic blood disease caused by unbalanced expression of peptide chains due to mutations in the β-globin gene, mostly caused by point mutations in the β-globin gene. β-thalassemia is one of the most common hereditary diseases in the world. According to statistics, more than 100 million people in the world carry the β-thalassemia gene. β-thalassemia is also one of the most common and most harmful genetic diseases in the southern provinces of my country. The carrier rate of β-thalassemia in some provinces of my country is shown in Table 1. Thalassemia genes are mainly point mutations, and β-thalassemia genes cover 46 point mutations in the southern Chinese population. [0003]...

AI Technical Summary

Problems solved by technology

The Sanger sequencing method is cumbersome to operate, which greatly limits its application in clinical diagnosis. The detection throughput of sequencing is also limited, and manual analysis is required for the detection results, which is time-consuming and labor-intensive.

RFLP is to carry out enzyme digestion reaction by identifying specific sequence sites. The method is simple and low-cost, but its limitation is that it can only detect limited mutations that can produce enzyme cutting sites. lead to false positive or false negative results

The result of reverse dot hybridization technology is interpreted by naked eyes, and the error rate is high, often resulting in repeated testing of a sample

ARMS-PCR needs to design corresponding primers for each mutation, and the amplification conditions of each pair of primers need to be optimized. If multiple mutations need to be detected at the same time, the operation is cumbersome, and false positive or false negative results may also occur

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