摘要
目的 应用脉冲场电泳 (PFGE)技术研究面肩肱型肌营养不良症 (FSHD)基因结构突变特征 ,并进行基因诊断。方法 研究对象包括 :110名健康对照 ,17例FSHD患者和 2 3名家系成员。低熔点胶包埋法抽提基因组DNA ,EcoRI、EcoRI/BlnI、XapI原位酶切 ,用脉冲电场凝胶电泳 (PFGE)分离 ,p13E 11探针Southern杂交 ,曲线拟合法计算分析片段的长度。结果 17例患者均存在 1条小于 35kb的 4 q35EcoRI致病片段 ;2 3名无症状家系成员中 ,发现 2例患者的母亲分别携带 2 5kb和 2 8kb的 4q35EcoRI短片段 ,因年龄较大 ,考虑为女性无症状顿挫型患者。另 2例患者的女性同胞分别携带 2 1 5kb和 31kb的 4 q35EcoRI短片段 ,因年龄较小 ,考虑为症状前患者。家族型和散发型患者均存在 4q→ 10 q型易位 ,各有 2例 ,其中 1例家族型患者为杂合易位 ;此外 ,在散发型患者中 ,发现 2例男性患者存在体细胞嵌合现象。结论 我国FSHD患者存在易位、杂合、体细胞嵌合等复杂现象。应用PFGE技术对FSHD患者进行基因诊断具有准确性和可靠性。
Objective Using pulse field gel electrophoresis (PFGE) to investigate the characteristics of gene structure and develop the gene diagnosis of facioscapulohumeral muscular dystrophy (FSHD) in the Chinese population Methods Totally 110 unrelated healthy individuals and 12 unrelated families including 17 affected and 23 unaffected members were studied Genomic DNA was extracted from peripheral blood lymphocytes according to the specific procedure designed to minimize DNA shearing, then cleaved with EcoRI, EcoRI/BlnI or XapI The cleaved DNA was separated by PFGE and Southern blotted with the probe p13E 11 The size of each fragment was calculated by “curve fitting” Results Each patient was detected carrying a less than 35 kb 4q derived EcoRI fragment Two female gene carriers of forme fruste,whose short 4q EcoRI fragment was 25 kb and 28 kb, and two presymptomatic patients,whose short 4q EcoRI fragment was 21 5 kb and 31 kb, were found in 23 clinically unaffected members A 4q derived EcoRI fragment translocating on chromosome 10 was found in two sporadic patients and two familial patients, one of them was heterogeneous In addition, somatic mosaicism was detected in 2 male sporadic cases Conclusion FSHD associated rearrangements including translocation, hybridization and somatic mosaicism were presented in Chinese FSHD patients PFGE for FSHD gene diagnosis should be accurate and reliable
出处
《中华神经科杂志》
CAS
CSCD
北大核心
2004年第6期521-525,共5页
Chinese Journal of Neurology
基金
国家自然科学基金资助项目 (3 0 0 0 0 182 )
福建省自然科学基金重点资助项目 (C992 0 0 0 2 )
福建省科技重大资助项目(2 0 0 2Y0 0 1)
关键词
脉冲场电泳技术
面肩肱型肌营养不良症
基因突变特征
基因诊断
Muscular dystrophy, facioscapulohumeral
Electrophoresis, gel, fulsed field
Translocation (genetics)
Mosaicism
