Duchenne형 근이영양증의 진단에 관한 연구

황기은(GE Hwang),조율희(YH Cho),심성한(SH Shim),최부영(BY Choi),차병헌(BH Cha),황윤영(YY Hwang),정성노(SR Chung),최수경(SK Choi),김영조(YJ Kim) 대한산부인과학회 1998 Obstetrics & Gynecology Science Vol.41 No.10

Dystrophin associated muscular dystrophies [dystrophinopathies] range from the severe Duchenne to the milder Becker muscular dystrophy [DMD and BMD]. Mapping and molecular genetic studies indicate that both are the result of mutations in the huge gene that encodes dystrophin. Approximately two-thirds of the mutations in both forms are deletions of one or many exons in the dystrophin gene. This study was performed in order to establish an diagnostic strategy for dystrophinopathy. In this study muscle biopsies were taken from the clinically suspected DMD and BMD patients, and immunohistochemical staining and Western blotting with anti-dystrophin antibodies were applied to the biopsy specimens. Also, the deletion analyses with multiplex PCRs for the 18 deletion prone exons, and linkage analyses for the diagnosis of carrier status and prenatal diagnosis were performed. We suggest that the following diagnostic strategy would be useful for the genetic counseling on dystrophinpathy families. 1. First of all clinically suspected dystrophinopathy patient should be subjected to deletion analysis with multiplex PCRs. If any deletions were found, the confirmatory diagnosis and prenatal diagnosis is possible. But the carrier diagnosis should be made with linkage analysis. 2. In the cases with no deletions, a muscle biopsy should be taken from the patient and protein study with immunohistochemistry or Western blotting should be performed. With normal dystrophin pattern, consider other neuromuscular diseases other than dystrophinopathy. If dystrophin is of reduced or increased size, with or without reduction in the amount of dystrophin, BMD should be suspected. If dystrophin is absent, DMD should be suspected. In this case prenatal and carrier diagnosis should be performed with linkage analysis.

Uncleaved Dystrophin Induce Cardiac Myocyte Apoptosis in Coxsackievirus Infected Balb/C Background Mice Heart

Park, Jin-Ho,Lee, Hye-Sun,Lee, Yun-Gyeong,Lim, Byung-Kwan 대한미생물학회 2014 Journal of Bacteriology and Virology Vol.44 No.3

It has been previously demonstrated that dystrophin is cleaved in the cardiac myocyte by the viral protease 2A following infection with Coxsackievirus B3 (CVB3). The viral protease 2A mediated cardiomyopathy can be prevented by inhibiting cleavage of dystrophin. However, it is less clear whether uncleaved dysdrophin have other heart protective effect in coxsackievirus infection. To address this, we generated a Balb/C background mouse that had a point mutation in dystrophin that prevents cleavage by protease 2A (KI). We show here that when mice expressing cleavage-resistant dystrophin were infected with CVB3, there was increased cardiac myocyte apoptosis. Bax and Bcl-$X_L$ mRNA ratio was significantly increased in KI mice heart compare to wild type mice heart. We found cleavage-resistant dystrophin induced the apoptosis related enzyme capspase-3 and caspase-8 activity. In addition, TUNEL stain was observed many TUNEL positive cardiac myocyte in KI mice heart compare to wild type mice heart (3.7% vs 0.3%). However, zVAD treatment for apoptosis blocking was significantly decreased myocardium damage and fibrosis in KI mice heart. These findings indicated that uncleaved dystrophin may have a critical role in cardiac myocyte viral propagation. Uncleaved dystrophin mutant induced cardiac myocyte apoptosis. It delayed coxsackievirus propagation in cardiac myocyte and could prevent cardiac myocyte death.

Uncleaved Dystrophin Induce Cardiac Myocyte Apoptosis in Coxsackievirus Infected Balb/C Background Mice Heart

박진호,이혜선,이윤경,임병관 대한미생물학회 2014 Journal of Bacteriology and Virology Vol.44 No.3

It has been previously demonstrated that dystrophin is cleaved in the cardiac myocyte by the viral protease 2Afollowing infection with Coxsackievirus B3 (CVB3). The viral protease 2A mediated cardiomyopathy can be preventedby inhibiting cleavage of dystrophin. However, it is less clear whether uncleaved dysdrophin have other heart protectiveeffect in coxsackievirus infection. To address this, we generated a Balb/C background mouse that had a point mutationin dystrophin that prevents cleavage by protease 2A (KI). We show here that when mice expressing cleavage-resistantdystrophin were infected with CVB3, there was increased cardiac myocyte apoptosis. Bax and Bcl-XL mRNA ratio wassignificantly increased in KI mice heart compare to wild type mice heart. We found cleavage-resistant dystrophin inducedthe apoptosis related enzyme capspase-3 and caspase-8 activity. In addition, TUNEL stain was observed many TUNELpositive cardiac myocyte in KI mice heart compare to wild type mice heart (3.7% vs 0.3%). However, zVAD treatmentfor apoptosis blocking was significantly decreased myocardium damage and fibrosis in KI mice heart. These findingsindicated that uncleaved dystrophin may have a critical role in cardiac myocyte viral propagation. Uncleaved dystrophinmutant induced cardiac myocyte apoptosis. It delayed coxsackievirus propagation in cardiac myocyte and could preventcardiac myocyte death.

Astroglial loss and edema formation in the rat piriform cortex and hippocampus following pilocarpine-induced status epilepticus

Kim, Ji-Eun,Yeo, Seong-Il,Ryu, Hea Jin,Kim, Min-Ju,Kim, Duk-Soo,Jo, Seung-Mook,Kang, Tae-Cheon Wiley Subscription Services, Inc., A Wiley Company 2010 Journal of comparative neurology Vol.518 No.22

<P>In the present study we analyzed aquaporin-4 (AQP4) immunoreactivity in the piriform cortex (PC) and the hippocampus of pilocarpine-induced rat epilepsy model to elucidate the roles of AQP4 in brain edema following status epilepticus (SE). In non-SE-induced animals, AQP4 immunoreactivity was diffusely detected in the PC and the hippocampus. AQP4 immunoreactivity was mainly observed in the endfeet of astrocytes. Following SE the AQP4-deleted area was clearly detected in the PC, not in the hippocampus. Decreases in dystrophin and α-syntrophin immunoreactivities were followed by reduction in AQP4 immunoreactivity. These alterations were accompanied by the development of vasogenic edema and the astroglial loss in the PC. In addition, acetazolamide (an AQP4 inhibitor) treatment exacerbated vasogenic edema and astroglial loss both in the PC and in the hippocampus. These findings suggest that SE may induce impairments of astroglial AQP4 functions via disruption of the dystrophin/α-syntrophin complex that worsen vasogenic edema. Subsequently, vasogenic edema results in extensive astroglial loss that may aggravate vasogenic edema. J. Comp. Neurol. 518:4612–4628, 2010. © 2010 Wiley-Liss, Inc.</P>

Duchenne/Becker 근이영양증에서의 Dystrophin 유전자 분석

박수연(Su Yeon Park) 고경남(Kyung Nam Koh) 임병찬(Byung Chan Lim) 강호석(Ho Seok Kang) 이경연(Kyoung Yeon Lee) 황희(Hee Hwang) 채종희(Jong Hee Chae) 최지은(Ji Eun Choi) 김기중(Ki Joong Kim) 황용승(Yong Seun Hwang) 대한소아신경학회 2004 대한소아신경학회지 Vol.12 No.1

목적 : 국내 Duchenne/Becker 근이영증에서의 dystrophin 유전자 결실은 빈도 및 그 분포를 알아보고 유전형과 임상형의 관계를 분석하여, 국내 유전 역학 자료를 마련하고 좀 더 효과적인 유전 상담 및 예방 방법을 모색해보고자 하였다. 방법 : 1999년 1월부터 2003년 7월까지 서을대학교병원에서 상기 질환으로 진단 후 추적 관찰중인 89명의 환자를 대상으로 하였다. 임상 진단 후 근생검을 시행, dystrophin 면역조직화학 염색을 하였으며 dystrophin 유전자 결실은 혈액에서 추출한 DNA를 이용, multiplex PCR 방법을 이용하여 진단하였다. 결과 : 국내 DMD/BMD 환자의 dystrophin 유전자 결실률은 54%였고, 결실 부위의 hot spot은 exon 44-54 부위로 전체 결실 환자의 80%를 차지하였다. 결실이 확인되지 않았던 22명의 환자에서 23개의 exon을 대상으로 직접염기서열 분석을 시행한 결과 6명의 환자에서 6종류의 점돌연변이를 발견하였다. 유전형과 임상형의 관계에 대한 분석 결과 중심형 결실이 근위형에 비해, 다중 결실이 단일 결실에 비해 증상 발생 연령이 어리고 진행이 다소 빠른 경향을 보였으나 통계적으로 유의한 관련성은 찾을 수 없었다. 결론 : 국내 DMD/BMD 환자의 결실률은 54%였으며 hot spot은 exon 44-54였다. 따라서 혈액 유전자 검사로 진단할 수 없는 약 50%의 환자의 진단을 위해서는 근생검 및 dystrophin 항체 면역 조직화학 검사를 통한 진단이 시행되어야 할 것이다. 또한 세밀한 가족력 조사를 포함하여 적절한 보인자 검사를 통해 합리적인 유전 강담을 진행하고 이에 기초한 질병 발생 예방 및 교육 등이 필요할 것으로 생각된다. Purpose : Duchenne/Becker muscular dystrophy(DMD/BMD) is an X-linked recessive disorder caused by mutations of dystrophin genes. The purpose of the present study is to determine the frequency and the patterns of dystrophin gene deletions and to investigate the correlation of genotypes and phenotypes. Methods : There were included a total of 89 children(88 boys and I girl) diagnosed as DMD/BMD by immunohistochemistry and/or genetic analysis from 1999 to 2003 at Seoul National University Children´s Hospital. We analyzed the genomic DNA by multiplex PCR using a 26 dystrophin exon primer set. Direct sequencing was performed on 23 exons(in which point mutations were detected in other previous reports) in 22 patients without deletions. Phenotype and genotype relationship analysis was performed on the basis of retrospective clinical reviews. Results : The frequency of dysmorphin gene deletions was 54%(32/59), which is lower than that of European and American data. Exon deletions were detected in 59 cases and the deletion "hot spots" were exon 44-54 constituting 80% of all deletions. In 6 cases without detectable deletions, 6 point mutations(3 nonsense mutations and 3 nucleotide variants) were detected The patients whose deletions were in the central parts or the patients with multiple exon deletions tended to show earlier symptom onsets and more rapid progressions of weakness but there were no statistical significances. Conclusion : Since deletions in dystrophin genes were detected in about 50% of the patients, studies on dystrophin protein expressions using muscle biopsy samples must be done for correct diagnosis.

Multiplex Ligation-dependent Probe Amplification (MLPA) 방법에 의한 디스트로핀 유전자 돌연변이 분자학적 진단의 유용성

조한나,홍지만,이경아,최영철 대한신경과학회 2010 대한신경과학회지 Vol.28 No.1

Background: Duchenne/Becker muscular dystrophy (DMD/BMD), which is the most common X-linked muscular dystrophy, is caused by mutations in the dystrophin gene. These mutations comprise deletions in approximately 55~65% of patients, duplications in 5~10%, and point mutations or small insertion/deletions in the remainder. Unfortunately, current diagnostic assays for dystrophin do not accurately detect duplication mutations or female carriers. In this study we employed multiplex ligation-dependent probe amplification (MLPA) analysis to detect deletions or duplications of the dystrophin gene in patients with DMD/BMD, and in potential female carriers. Methods: A total of 41 subjects was recruited for this study, comprising 35 male DMD/BMD patients, 1 female patient with Turner syndrome, and 5 females with a family history of DMD/BMD. The MLPA method was employed to determine the copy number of each of the 79 exons of the dystrophin gene in the 41 subjects. Results: MLPA analysis for dystrophin was informative in 71.4% (25/35) of patients with DMD/BMD patients, identifying deletions in 60.0% (21/35) and duplications in 11.4% (4/35). MLPA analysis showed the presence of a deletion of the DMD gene in one female patient with Turner syndrome. Of the five female patients with a family history of DMD/BMD, this assay revealed exon deletion in one and duplications in one. Conclusions: The reported findings reveal that the MLPA method is a powerful tool for detecting duplications and female carriers, as well as DMD gene deletions. MLPA should be considered the method of choice for an initial genetic analysis of DMD/BMD patients. Background: Duchenne/Becker muscular dystrophy (DMD/BMD), which is the most common X-linked muscular dystrophy, is caused by mutations in the dystrophin gene. These mutations comprise deletions in approximately 55~65% of patients, duplications in 5~10%, and point mutations or small insertion/deletions in the remainder. Unfortunately, current diagnostic assays for dystrophin do not accurately detect duplication mutations or female carriers. In this study we employed multiplex ligation-dependent probe amplification (MLPA) analysis to detect deletions or duplications of the dystrophin gene in patients with DMD/BMD, and in potential female carriers. Methods: A total of 41 subjects was recruited for this study, comprising 35 male DMD/BMD patients, 1 female patient with Turner syndrome, and 5 females with a family history of DMD/BMD. The MLPA method was employed to determine the copy number of each of the 79 exons of the dystrophin gene in the 41 subjects. Results: MLPA analysis for dystrophin was informative in 71.4% (25/35) of patients with DMD/BMD patients, identifying deletions in 60.0% (21/35) and duplications in 11.4% (4/35). MLPA analysis showed the presence of a deletion of the DMD gene in one female patient with Turner syndrome. Of the five female patients with a family history of DMD/BMD, this assay revealed exon deletion in one and duplications in one. Conclusions: The reported findings reveal that the MLPA method is a powerful tool for detecting duplications and female carriers, as well as DMD gene deletions. MLPA should be considered the method of choice for an initial genetic analysis of DMD/BMD patients.

Duchenne/Becker Muscular Dystrophy의 진단

이지훈(Jee Hun Lee),이란(Ran Lee),권영세(Young Se Kwon),이종화(Jong Hwa Lee),채종희(Jong Hee Chae),김기중(Ki Joong Kim),주세익(Saeick Joo),박성섭(Sungsup Park),조한익(Han Ik Cho),지제근(Jegeun Chi),황용승(Yong Seung Hwang) 대한소아신경학회 2000 대한소아신경학회지 Vol.8 No.2

목적: Duchenne/Becker muscular dystrophy(DMD/BMD)는 X-연관 열성유전하는 진행성 근육질환으로 dystrophin 유전자의 돌연변이에 의하여 발병한다. 유전자 이상은 55-65%의 환자에서 결실에 의하여 일어나며, 최근에는 이에 근거한 분자유전학적인 방법을 진단에 이용하고 있다. 저자들은 근이양증 환자들의 임상양상을 정리하고, dystrophin 유전자 검사 및 면역조직화학 검사의 양상을 분석하며, 이에 근거한 국내 환자들의 분자 유전학적 및 형태학적 진단의 표준을 제시하고자 본 연구를 시행하였다. 방법: 1989년부터 1999년까지 서울대학교병원 소아과에서 임상양상, 혈청 creatine kinase, 근전도 및 근육생검 소견에 근거하여 근이양증으로 진단된 환아 69명을 대상으로 후향적 연구를 하였다. 34명에서 26 set의 primer를 이용한 multiplex PCR을 통한 유전자 결실 검사를 시행하였으며, 5명에서 dystrophin 면역조직화학 염색을 시행하였다. 결과: 대상 환자의 내원시 평균연령은 5년 8개월이었으며, 평균 초발연령은 3년 6개월이었다. 첫 증상은 하지의 근력약화, 우연히 발견된 간효소의 상승, 보행이상, 운동발달 지연 등이었다. 41%에서 운동발달 지연이 있었고, 42%는 15개월 이후에야 혼자 걷기가 가능하였다. 대부분에서 비복근의 가성비대가 관찰되었으며, 혈청 creatine kimase는 평균 11,232IU/L였고, 44%에서 심전도상 이상소견을 보였다. 근육생검을 시행한 63례 모두에서 전형적인 병리소견이 관찰되었다. 면역조직화학 염색을 실시한 5명 중 2례에서 DYS1, 2, 3 전체에서 염색이 되지 않았고, 3례에서는 DYS1, 2에서 불규칙한 염색상을 보이고, DYS3에서는 염색이 되지 않았다. Multiplex PCR을 시행한 34명 중 14례(41%)에서 결실이 확인되었으며, 이 중 11례 (78.6%)에서 hot spot으로 알려진 exon 44와 exon 55 사이의 결실이 있었다. 결론: Multiplex PCR을 이용한 dystrophin 유전자 검사는 전체 환자의 50% 정도만을 진단할 수 있으므로 나머지 환자의 진단을 위해서는 dystrophin 단백의 발현에 대한 검사가 같이 시행되어야 할 것이다. 분자유전학적인 진단이 DMD/BMD의 선별진단 도구가 되고 있으므로, 근육생검의 침습성을 감안하면, 임삭적으로 의심이 되는 경우 유전자의 흔한 돌연변이 유무를 우선적으로 확인한 후, 결손이 없는 경우 근육생검을 실시하여 광학현미경적 소견과 아울러 면역조직화학적 소견을 확인하여 다른 근이영양증과의 감별을 시도하는 것이 가장 합리적인 진단과정으로 생각된다. Purpose: Duchenne/Becker muscular dystrophy(DMD/BMD) is an X-linked recessive disease caused by the mutation of dystrophin gene. Since the majority of mutations are deletions, recent diagnosis is made by the moleculargenetic tools. The authors summarized the clinical characteristics, and analyzed the moleculargenetic and immunohistochemical characteristics of DMD/BMD. Methods: We reviewed the clinical and laboratory findings of 69 patients diagnosed as DMD/BMD from 1989 to 2000. Multiplex PCR using 26 primer sets was performed on 34 cases, and immunohistochemical staining using dystrophin antibody was done on 5 cases. Mutation profile and phenotype-genotype relationship were analyzed. Results: 1) Mean age of onset was 3 years and 6 months. The presenting symptoms were motor weakness of the lower extremities, incidentally found elevated hepatic enzyme level, abnormal gait and motor developmental delay. Forty one percent had history of motor developmental delay, and most patients showed pseudohypertrophic calf muscles. Mean serum creatine kinase level was 11,232IU/L, and 44% revealed abnormal electrocardiogram. 2) All of the 63 cases showed typical histological findings of muscular dystrophy. Of the 5 cases with immunohistochemical staining, 2 showed complete(DYS1, 2 and 3) and 3 showed partial(DYS3) absence pattern. 3) Of the 34 cases on which multiplex PCR was performed, 14 showed deletions, and 11 of them had deletions between exon 44 and 55. Conclusion: Since the deletions were detected in less than 50% of the patients with multiplex PCR, tools for dystrophin protein expression must be combined for the correct diagnosis. Considering the invasiveness of muscle biopsy, we conclude immunohistochemistry should be followed in the cases with negative results in multiplex PCR, although moleculargenetic study is the primary diagnostic tool.

심근침범을 동반한 Becker형 근이영양증 환자에서 확인한 Dystrophin 유전자 Exon결실

김광일,오병희,이무용,채인호,신수,박성섭,김효수,손대원,이명묵,박영배,최윤식,이영우 대한순환기학회 1998 Korean Circulation Journal Vol.28 No.5

Frameshift Deletion Mechanisms in Egyptian Duchenne and Becker Muscular Dystrophy Families

Nasser Attia Elhawary 한국분자세포생물학회 2004 Molecules and cells Vol.18 No.2

Partial gene deletion is the major type of mutation leading to Duchenne muscular dystrophy (DMD) and its mild allelic form, Becker muscular dystrophy (BMD). Amplification of the genomic DNAs of 152 unrelated dystrophin patients using multiple primers detected 78 (51.3%) probands with deletion mutations. We predicted the translational reading frame for all the deletions in Egyptian dystrophin males. The frameshift rule was confirmed positively ranging for 50 to 67% of the cases depending on the type of disease. We discuss ways of accounting for some exceptions from the frameshift hypothesis in the central and proximal regions. These explanations may help in developing procedures for reducing the severity of dystrophin phenotypes to restore the correct frame by disrupting the translational fidelity. Great efforts have been put into the development of effective ‘gene correction’ procedures via such intrinsic mechanisms. In addition, we mapped regional difference in deletion mutation frequencies within the DMD gene locus between the different Egyptian governorates. There were no double deletions in the Egyptian dystrophin males.

Duchenne/Becker 근이영양증에서의 임상적, 면역조직학적 및 유전학적 소견

서창덕(Chang-Deok Seo),이윤정(Yoon-Jung Lee),정민희(Min-Hee Jeong),이은혜(Eun-Hye Lee),염미선(Mi-Sun Yum),고정민(ung-Min Ko),유한욱(Han-Wook Yoo),고태성(Tae-Sung Ko) 대한소아신경학회 2009 대한소아신경학회지 Vol.17 No.1

목 적:본원에서 Duchenne 및 Becker 근이영양증(DMD/BMD)으로 진단된 환자에서 임상양상과 multiplex PCR 또는 MLPA를 이용한 디스트로핀 유전자 결손, 면역조직화학 염색에 의한 디스트로핀 발현을 조사하여 각 임상형의 특징과 각 검사의 진단적 유용성에 대해 고찰해 보았다. 방 법:1989년 6월부터 2008년 12월까지 서울아산병원에서 진단시 혈청 Creatinine Kinase(CK) 증가가 있고 근육질환의 임상소견을 보이는 환자에서 조직학적 소견 또는 디스트로핀 유전자 검사소견이 DMD/BMD에 부합되는 93명을 대상으로 의무기록을 후향적으로 분석하였다. 성별, 나이, 첫 임상증상의 종류와 발병 연령, 진단시 Gower sign 유무, 가족력, 진단시 혈청 CK 수치, 근전도 및 신경전달속도검사, 대퇴근의 근육생검 및 면역조직학적 검사, 말초혈액을 이용한 디스트로핀 유전자 검사(multiplex PCR 또는 MLPA)를 조사하였다. 또한 frame 유지여부와 임상표현형의 일치율을 구해 보았다. 결과:DMD 58명, BMD 13명, 비분류군(unclassified) 19명, DMD/BMD carrier 3명이었다. 증상이 처음 발견된 연령은 평균 4.7±3.2세(1-17세)였고 첫 증상은 보행장애가 46명(49%)으로 가장 많았고 간효소 상승이 우연히 발견된 경우가 35명(37 %)으로 두번째로 많았다. 진단시 혈청 CK는 평균 14,758±11,792(633-6,1349) IU/L이었고 DMD에서는 18,402±12,117(633-61,349) IU/L, BMD에서는 6,675±6,414(814-17,898) IU/L, unclssified에서는 11,103±9,032(2,623-39,549)IU/L, PMD carrier는 1,281±637(718-1,974),IU/L이었다. 근육 생검을 한 75명중 clone 13H6(N-terminus)를 사용하여 DMD 48명과 BMD 10명에서 면역조직화학염색을 시행하였는데 DMD에서는 48명 모두 디스트로핀이 발현되지 않았으며 BMD에서는 10명 모두 불완전/부분적으로 발현되었다. Multiplex PCR을 시행한 54명중 28명(51%)에서 결손이 발견되었고 MLPA를 시행한 14명 중 13명(92%)에서 결손이 발견되었고 이중 2명은 이형접합결손이었다. Frame 유지여부와 임상표현형의 일치율은 95%를 보였다. 결론:multiplex PCR를 시행한 10명의 환자와 MLPA를 시행한 11명의 환자를 대상으로 frame 유지여부와 임상표현형과의 일치율을 구해보았는데 일치율은 95%로 높았다. 대상군이 적어 추가적인 연구가 필요할 것으로 보이지만 MLPA를 통해 in frame과 out of frame을 결정한 후 DMD/BMD를 예상하는 것은 근육생검을 시행하지 않는 환자에 있어서는 임상적으로 유용하겠다. 또한 MLPA를 통한 carrier의 진단이 유전상담에 있어 큰 도움이 되겠다. Purpose:This retrospective study was designed to know the relation between clinical features, genetics, and immunostaining findings among children with Duchenne muscular dystrophy (DMD)/Becker muscular dystrophy (BMD) and the validity of the diagnostic tools for muscular dystrophy. Methods:The medical records and computerized databases of 93 patients diagnosed with DMD/BMD from June 1989 to December 2008 were reviewed retrospectively. Demographic characteristics including clinical features, serum creatinine kinase(CK) level, electromyogram(EMG) and nerve conduction velocity(NCV), muscle biopsy, immunochemical staining for dystrophin, and the deletion of dystrophin gene were analyzed. We calculate the concordance rate between type of frame (in or out of frame) and phenotype. Results:58(62%) children were diagnosed with DMD, 13(14%) BMD, 19(20%) unclassified dystrophy, and 3(3%) DMD/BMD carriers. The mean age of symptom onset was 5.0±3.5 years(range, 1-17). 46(49%) children presented gait disturbance and 35(37%) elevation of liver enzymes. The mean value of serum CK enzyme was 14,758±11,792 IU/L (range, 633-61,349). There was no dystrophin in the immunochemical stain among 48 DMD children and at least partial or incomplete dystrophin among 10 BMD children. 28/54(51%) children had dystrophin gene deletion in multiplex PCR and 13/14(92%) in Multiplex Ligation-dependent Probe Amplification(MLPA). The loss of heterozygosity was shown in 2 children by MLPA. The overall concordance rate between type of frame(in or out of frame) and phenotype was 95% in this study. Conclusion:Despite of small population, this finding indicates that the determination of type of frame (in or out of frame) by MLPA may be helpful in differential diagnosis of DMD/BMD. In addition, we surmise that the detection of carrier by MLPA is helpful in genetic counseling.