Multiple PCR primers, kit and application

Abstract

The invention provides multiple PCR primers for detecting beta-thalassemia mutation based on next-generation sequencing. The multiple PCR primers are two or more primer pairs selected from SEQ ID NO:1 and SEQ ID NO:2 primer pair, SEQ ID NO:3 and SEQ ID NO:4 primer pair, SEQ ID NO:5 and SEQ ID NO:6 primer pair, SEQ ID NO:7 and SEQ ID NO:8 primer pair, SEQ ID NO:9 and SEQ ID NO:10 primer pair, and SEQ ID NO:11 and SEQ ID NO:12 primer pair.

Description

[0001] related application [0002] This application is a divisional case of Chinese patent application 201510740909.5 with an application date of November 4, 2015, and titled "Multiple PCR primers and methods and applications for detecting β-thalassemia mutations based on next-generation sequencing technology". technical field [0003] The invention belongs to the field of molecular biology and relates to primers, in particular to multiple PCR primers, kits and applications. Background technique [0004] β-thalassemia is a monogenic genetic blood disease caused by unbalanced expression of peptide chains due to mutations in the β-globin gene (HBB), 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 ...

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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