Detection kit and method for Duchenne/Bell muscular dystrophy based on multiple PCR capture technology

Abstract

The invention discloses a multiplex PCR (Polymerase Chain Reaction) trapping primer group for a DMD (Duchenne Muscular Dystrophy) gene, a mutation detection kit for the DMD gene and a mutation detection method for the DMD gene. The inventors provide the multiplex PCR trapping primer aiming at the sequences of 79 exons and bilateral 50bp introns and 23 known intron pathogenic mutations of the DMD gene; moreover, the mutation detection method of the DMD gene is provided based on the primer group; the mutation detection method is high in detection sensitivity, good in accuracy, highly consistentin amplification efficiency and low in requirement on quality of a DNA (Deoxyribonucleic Acid) sample, and is used for solving the problem that the complete detection on the mutation information, particularly on loss of heterozygosity / repetition, of the DMD gene is difficultly carried out in one time by adopting a multiplex PCR trapping technique in the prior art.

Description

technical field [0001] The invention belongs to the field of gene sequencing, and in particular relates to a detection kit and a method for Duchenne / Bell muscular dystrophy based on multiplex PCR target sequence capture technology. Background technique [0002] Duchenne muscular dystrophy is a fatal X-linked recessive genetic disease, which can be divided into pseudohypertrophic muscular dystrophy (Duchenne muscular dystrophy, DMD) according to the degree of progress BMD (Becker muscular dystrophy, BMD), both are allelic clinically heterogeneous diseases. Among them, the main clinical features of DMD are progressive atrophy and weakness of the proximal skeletal muscles of the extremities, pseudohypertrophy of the gastrocnemius muscle of the calf, involving both the respiratory muscles and the myocardium, and some patients are accompanied by mental retardation. Loss of walking ability and death in the 20s occurs in up to 1 in 3500 male live births. Compared with DMD, the ag...

AI Technical Summary

Problems solved by technology

Due to the complexity of the DMD gene and the diversity of mutation types, traditional DMD gene mutation detection methods such as Southern hybridization (Southern blotting), denaturing high performance liquid chromatography (DHPLC), multiple junction-dependent Probe amplification (Multiplex ligation-dependent probe amplification, MLPA), microarray-comparative genomic hybridization (array comparative genomic hybridization, arrayCGH), etc., all have some outstanding problems, such as only partial mutation types and known mutations can be detected , the detection throughput is low, the detection cycle is long, and the detection accuracy is low; about 30% of the children need a second detection, which greatly increases the detection cycle and detection cost

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