Study On The Dystrophin Gene Of Duchenne/becker Muscular Dystrophy Patients And Carriers Applying MLPA

Objective:1,To investigate the value of multiplex ligation-dependent probe amplification (MLPA) method in detecting Dystrophin gene in Duchenne/Becker muscular dystrophy (DMD/BMD) patients, making the gene diagnosis system perfect.2,To analyze the correlation between the Genotype and phenotype of Duchenne/Becker muscular dystrophy, providing evidence for Prognostic analysis and gene therapy.3,To investigate the advantages of MLPA to detect Dystrophin gene in DMD/BMD carriers, providing reliable evidence for genetic counseling.Methods:1,Dystrophin gene was analyzed by MLPA in 355 patients with DMD/BMD and it were verified by PCR and sequencing for a single exon deletion or a low ratio of peak area of exon.2,SPSS 13.0 package was applied to analyzed the correlation between the frameshift mutations, mutation range, mutation type of Dystrophin gene and the phenotype of patients.3,Carrier was Detected by MLPA in 46 mothers of patient with deletion mutation and 8 mothers of patient with duplication mutation, the carrier incidences of two groups were compared.Results:1,187 cases were detected to have deletion of one or more Dystrophin exons, 34 patients had duplication mutation, 5 patients had point mutation, and no deletions or duplications could be detected in the remaining 129 patients.2,There is significant correlation between translational reading frame and the phenotype of patients, 91.55% DMD and 86.27% BMD follow the"frame rule", but there was no statistical significance between clinical phenotype and the mutation range or mutation type of Dystrophin gene.3,28 of the 46 mothers of patient with deletion mutation and 6 of the 8 mothers of patient with duplication mutation were identified as carrier, there was no statistical significance between the carrier incidences of the 2 groups.Conclusions:1,MLPA can detect the deletion and duplication mutation of all the 79 exons of Dystrophin gene, improving the efficiency, sensitivity and stability of genetic diagnosis, Detection of a single exon deletion or a low ratio of peak area of an exon by Combining PCR and sequencing can improve the accuracy of the results.2,The genotype–phenotype correlation in most of the DMD / BMD patients follow the"frame rule", providing evidence for gene therapy, but there was no correlation between clinical phenotype and the mutation range or mutation type of Dystrophin gene.3,MLPA can quantify the copy number of Dystrophin Gene accurately, being adequate for detecting carrier and providing reliable evidence for genetic counseling.