Detection of DMD by Multiplex Ligation-depenclent Probe / 医学研究杂志

ABSTRACT

Objective Duchenne muscular dystrophy(DMD)is one of the most common X-linked recessive neuromuscular degeneration diseases.It is caused by genetic defects of dystropin gene with deletion,duplication,or point mutation that results in clinical muscle fatigue and dystrophy.Usually,gene deletion of one or a few exons of dystrophin accounts for about 55%～65% patients,duplication for about 5%～10% patients and point mutation for 25%.Most of hot-spot deletion mutation of DMD can be detected by multiplex PCR and the point mutation can be detected by PCR/sequencing analysis,however,it remains a challenge to detect duplication.The recently developed MLPA(multiplex ligation-dependent probe amplification)is an efficient procedure that can accurately analyze the copy number and deletion mutation of whole dystropin gene.Methods A validation for simultaneous detection of entire dystropin gene was performed with two reactions.Both of which detect 39 and half exons of dystrophin gene.Results Nine out of 15 patients with DMD were found to have deletion mutation in different exons of dystrophin gene.Among these 9 patients,7 were found having deletion previously with multiplex PCR for mutation of hot-spot by Peking Union Medical University.Two patients who had not been found deletion by multiplex PCR were shown to have rare deletion at exon 18 or 43 in this study.Conclusions MLPA provides a simple,rapid and accurate method of simultaneously detecting homozygous,heterozygous deletions and duplication mutation in two single reactions for all exons of dystrophin gene,which may be applied into clinical molecular analysis for DMD.