CAJ | 학술논문

目的应用多重连接依赖性探针扩增(MLPA)技术对假肥大型肌营养不良(DMD及BMD)患者及其家系成员进行dystrophin基因分析,探讨MLPA定量技术在本病重复突变及携带者检测中的优势.方法以355例DMD及BMD患者、46名缺失型患者之母和8名重复型患者之母为研究对象,应用MLPA技术对dystrophin基因全长外显子进行分析,对于单一外显子缺失的样本采用PCR及测序进行验证.结果经MLPA分析,全部355例患者中190例为dystrophin基因缺失型患者,在其余非缺失型患者中检测出34例重复型突变.此外,在46名缺失型患者的母亲中发现了28名携带者,在8名重复型患者的母亲中发现了6名携带者,两组患者母亲携带者频率差异无统计学意义.经过测序验证,在1例单一外显子缺失的患者中发现17号外显子存在AGGGAACAGATCCTGGTAAAGCA小片段缺失.结论与传统的定量方法相比,MLPA定量技术可对DMD及BMD患者全长外显子区域同时进行缺失、重复分析,并能对患者家系成员的携带状态进行判定.此外,MLPA检测结果受模板DNA的浓度及纯度影响较小.
목적 응용다중련접의뢰성탐침확증(MLPA)기술대가비대형기영양불량(DMD급BMD)환자급기가계성원진행dystrophin기인분석,탐토MLPA정량기술재본병중복돌변급휴대자검측중적우세.방법 이355례DMD급BMD환자、46명결실형환자지모화8명중복형환자지모위연구대상,응용MLPA기술대dystrophin기인전장외현자진행분석,대우단일외현자결실적양본채용PCR급측서진행험증.결과 경MLPA분석,전부355례환자중190례위dystrophin기인결실형환자,재기여비결실형환자중검측출34례중복형돌변.차외,재46명결실형환자적모친중발현료28명휴대자,재8명중복형환자적모친중발현료6명휴대자,량조환자모친휴대자빈솔차이무통계학의의.경과측서험증,재1례단일외현자결실적환자중발현17호외현자존재AGGGAACAGATCCTGGTAAAGCA소편단결실.결론 여전통적정량방법상비,MLPA정량기술가대DMD급BMD환자전장외현자구역동시진행결실、중복분석,병능대환자가계성원적휴대상태진행판정.차외,MLPA검측결과수모판DNA적농도급순도영향교소.
Objective To analyze the dystrophin gene in patients with Duchenne/Becker muscular dystrophy (DMD/BMD) and their family members by multiplex ligation-dependent probe amplification (MLPA) method and to evaluate the application of this method in the mutations detection. Methods The whole dystrophin gene (79 exons) was analyzed by MLPA in 355 patients with DMD/BMD, the mothers of 46 patients with deletion mutation and the mothers of 8 patients with duplication mutation. The results were verified by PCR and sequencing when single exon deletion was found. Results One hundred and ninety cases were found to have deletion of one or more dystrophin exons, and 34 patients were identified to have duplication mutations. In 46 mothers of patients with deletion mutations, 28 were identified the mutations;and of 8 mothers of patients with duplication mutations, 6 were identified the mutations. There was no statistical significance between the carrier incidences in the 2 groups. A 23 bp deletion of "AGGGAACAGATCCTGGTAAAGCA" fragment in exon 17 was found in a patient. Conclusions Comparing with the traditional quantitative methods, MLPA can detect the deletion and duplication mutation in all the 79 exons of dystrophin gene in DMD/BMD patients, and can identify the carrier status in their family members. Furthermore, MLPA is not apt to be interfered by the concentration and purity of DNA template.