Multiple PCR for detecting the deletion of DMD gene exon in patients with Duchenne/Becker muscular dystrophy

Objective To develop a consistent and effective gene diagnostic method for Duchenne/Becker muscular dystrophy(DMD/BMD),so as to provide a clinical reference for the differential diagnosis of DMD/BMD diseases.Methods Clinical data of sixty-four male DMD/BMD patients,Han nationality,aged from 0.7 to 45(9.34±1.13)years,were collected between May 2008 and May 2010 from the Departments of Neurology,Pediatrics,and Gynaecology and Obstetrics of affiliated hospitals of Third Military Medical University.Genomic DNA was extracted from 5ml of peripheral blood of each patient.Determination of exon deletion in DMD gene of DMD/BMD patients was performed by amplifying 18 exon in hotspot deletion regions of DMD gene with multiple PCR(mPCR),and then the deletion type of DMD gene exon was confirmed.Results Deletions of varied exons in hotspot deletion regions of DMD gene were found in 31(48.4%)out of 64 patients with DMD/BMD,among which exon 50 deletion ranked the top(35.5%,11/31),followed by exon 47 and 43 deletion,accounting for 32.3%(10/31)and 25.8%(8/31),respectively.Exon 45 deletion was found in 6 out of the 31 patients(19.3%),while exon 49 deletion existed in same number of patients.Exon 19 deletion,as well as the 48 deletion,was respectively found in 5 out of the 31 patients(16.1%).In summary,the exon deletion happened mainly in the central and 5-end hotspot deletion region of DMD gene.Conclusions Multiple PCR,extensively used domestically and abroad,plays a vital role in assisting clinical diagnosis of DMD/BMD.It still is worthwhile to explore the other deletion hotpot,duplicated mutational hotpot and point mutation hotpot of DMD gene,and,based on them,new methods for DMD/BMD gene diagnosis can be established,thereby the diagnostic rate of DMD/BMD disease can be elevated.