Method and kit for detecting Duchenne muscular dystrophy gene defects, and primer composition

Abstract

The invention relates to a method and a kit for detecting Duchenne muscular dystrophy gene defects, and a primer composition. For the Duchenne muscular dystrophy gene defects, a pre-embryo implantation genetic detection method which is strong in universality, high in diagnosis rate and low in cost is established in a second-generation sequencing platform. According to the method, multiple PCR is carried out on a detected sample for a coding region locus and a target SNP locus of a Duchenne muscular dystrophy gene, and a library is built; then a multiplex PCR product is subjected to high-throughput sequencing analysis, and a haplotype of a mutant allele is constructed by combining with family information, so that a genotype of an embryo is judged; and finally a Duchenne muscular dystrophy gene defect detection result of an offspring is determined. According to the scheme, a cell level pre-experiment is omitted; and based on a high-throughput sequencing technology, different label sequences can be added to samples, so that a large number of samples can be analyzed at one time, and the average sequencing depth can reach more than 100X.

Description

technical field [0001] The invention relates to the field of gene detection, in particular to a detection method, a primer composition and a kit for Duchenne muscular dystrophy gene defects. Background technique [0002] Duchenne muscular dystrophy (DMD) is a common X-linked recessive genetic fatal neuromuscular disease with an incidence of about 1 / 3500 live-born male babies, with no significant geographical or racial differences. Patients are usually male and the onset is often in childhood. The main clinical manifestations are progressive weakness and atrophy of the skeletal muscles of the whole body, pseudohypertrophy of the gastrocnemius muscle of the calf, abnormal walking posture, easy to fall, difficulty climbing stairs and squatting up, slowly progressive aggravation of the disease, and gradually bedridden, often due to respiratory failure at the age of 20 and / or death from cardiac insufficiency. The DMD pathogenic gene is the largest human gene discovered so far, ...

AI Technical Summary

Problems solved by technology

However, karyomapping cannot detect gene mutations, so there must be samples of probands or suitable family members. If there are not enough family samples, additional mutation detection is required.

Although there are some regions with relatively high incidence of mutations in DMD pathogenic genes, all exons are reported to have mutations, and 30% of patients have new mutations. The DMD gene is as long as 2.4Mb, and the recombination probability within the gene is about 12 %

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