Amplifying system and reagent kit for detecting DMD (Duchenne muscular dystrophy) gene exon copy number variation

Abstract

The invention relates to an amplifying system for detecting human DMD (Duchenne muscular dystrophy) gene exon copy number variation. The amplifying system comprises an amplifying composition corresponding to the detection of 10 groups of exon copy number variation, wherein the amplifying composition comprises oligonucleotide sequences as shown from SEQID No.1 to SEQID No.36; the 10 exon purpose fragments including human DMD gene exons 4,8,17,44,45,47,48,50,51,52 are amplified through a real-time multiple fluorescent quantitation PCR reaction; and according to the detection result, whether thehuman DMD gene exon is subjected to deletion mutation or repeat mutation is judged. The invention also discloses a reagent kit for detecting human DMD (Duchenne muscular dystrophy) gene exon copy number variation. The amplifying system disclosed by the invention has the beneficial effects that through the detection result, the distinguishing of DMD carriers from DMD patients and normal people canbe realized, the result is reliable, the repeatability is good, the operation is easy, the equipment requirement is low, and the reagent cost is low; and the amplifying system can cover about 90% of copy number variation in the DMD carriers or the DMD patients, so that good balance between the detection cover degree and the detection exon number is achieved.

Description

technical field [0001] The invention relates to the field of gene molecular detection, in particular to an amplification system and a kit for detecting the variation of the exon copy number of the DMD gene. Background technique [0002] Duchenne / Beckman muscular dystrophy (DMD / BMD) is a group of genetic diseases caused by DMD gene mutation leading to dystrophin protein deficiency. Clinical phenotypes include: asymptomatic hypercreatine kinaseemia, muscle spasm / myalgia, quadriceps myopathy, X-linked dilated cardiomyopathy (X-linked dilated cardiomyopathy, XLDCM), etc. The incidence of DMD in male infants is about 1 / 3500-1 / 4000, and the onset is more than 5-6 years old. It manifests as slow walking, easy to fall, and abnormal walking posture. Died of cardiopulmonary failure. The incidence of BMD in males is about 1 / 8000-1 / 10000. The clinical symptoms appear later than DMD, and the ability to walk can be maintained until the age of 16. The disease progresses relatively slowly...

AI Technical Summary

Problems solved by technology

[0009] The MLPA detection technology of DMD gene has good accuracy, but the reagent cost of MLPA detection technology is relatively high, and the capillary electrophoresis equipment used for fragment screening, such as ABI 3130 or 3500, is expensive

In addition, there are many links in the MLPA detection technical process, mainly including four main steps of overnight hybridization, probe ligation, PCR amplification, and fragment screening. Many links put forward higher requirements for the experimental skills and proficiency of operators , which also increases the possibility of large fluctuations in the RQ value and errors in the sample number

In addition, the most valuable application of DMD gene copy number detection is the screening of female carriers. In this way, large sample size detection is required. Due to its many technical links, MLPA detection technology has certain advantages in dealing with large sample size detection. practical difficulty of

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