Mutational scanning of the ABCR gene with double-gradient denaturing-gradient gel electrophoresis (DG-DGGE) in Italian Stargardt disease patients
Mutational scanning of the ABCR gene with double-gradient denaturing-gradient gel electrophoresis (DG-DGGE) in Italian Stargardt disease patients
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DOI：
10.1007/s004390100583
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发表时间：
2001-09-01
期刊：
影响因子：
5.3
通讯作者：
Cremonesi, L
中科院分区：
文献类型：
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作者：
Fumagalli, A;Ferrari, M;Cremonesi, L
视网膜特异性ABC转运蛋白（ABCR）基因突变导致常染色体隐性遗传性Stargardt病（arSTGD）。在先前发表的研究中，arSTGD患者中ABCR的突变检测效率在30%至66%之间，这是由于所采用方法的高等位基因异质性和技术限制。建立了双梯度变性梯度凝胶电泳筛选ABCR基因的条件。通过分析具有先前表征的ABCR突变的DNA样品来评估该方法的功效。该方法应用于44名意大利arSTGD患者的突变检测，对应于36个独立的基因组，以评估该种族组中ABCR突变的性质和频率。在34/36例（94.4%）STGD患者中，确定了37个序列变化，包括26个错义，6个移码，3个剪接和2个无义变异。其中，20个以前没有描述过。在受影响的个人和匹配的对照中检测到几个多态性。我们的研究结果扩展了STGD患者中发现的突变谱，并表明存在意大利人群特有的分子缺陷子集。在四个健康对照个体中鉴定出至少两个疾病相关突变，表明一般人群中变异ABCR等位基因的携带频率高于预期。在我们的系列基因型-表型分析表明，一些突变的性质和位置与STGD疾病的特定检眼镜特征之间可能存在相关性。
Mutations in the retina-specific ABC transporter (ABCR) gene are responsible for autosomal recessive Stargardt disease (arSTGD). Mutation detection efficiency in ABCR in arSTGD patients ranges between 30% and 66% in previously published studies, because of high allelic heterogeneity and technical limitations of the employed methods. Conditions were developed to screen the ABCR gene by double-gradient denaturing-gradient gel electrophoresis. The efficacy of this method was evaluated by analysis of DNA samples with previously characterized ABCR mutations. This approach was applied to mutation detection in 44 Italian arSTGD patients corresponding to 36 independent genomes, in order to assess the nature and frequency of the ABCR mutations in this ethnic group. In 34 of 36 (94.4%) STGD patients, 37 sequence changes were identified, including 26 missense, six frameshift, three splicing, and two nonsense variations. Among these, 20 had not been previously described. Several polymorphisms were detected in affected individuals and in matched controls. Our findings extend the spectrum of mutations identified in STGD patients and suggest the existence of a subset of molecular defects specific to the Italian population. The identification of at least two disease-associated mutations in four healthy control individuals indicates a higher than expected carrier frequency of variant ABCR alleles in the general population. Genotype-phenotype analysis in our series showed a possible correlation between the nature and location of some mutations and specific ophthalmoscopic features of STGD disease.