DNA sequencing errors in molecular diagnostics of filamin myopathy
DNA sequencing errors in molecular diagnostics of filamin myopathy
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DOI：
10.1515/cclm.2010.272
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发表时间：
2010-10-01
影响因子：
6.8
通讯作者：
Goldfarb, Lev G.
中科院分区：
文献类型：
--
作者：
Odgerel, Zagaa;van der Ven, Peter F. M.;Goldfarb, Lev G.
背景资料：细丝蛋白肌病是一种神经肌肉疾病，主要表现为肢带肌无力，许多患者患有膈肌麻痹和心肌病，由细丝蛋白C（FLNC）基因突变引起。目的：探讨细丝蛋白肌病的分子诊断方法。方法：采用分子克隆、RT-PCR和real-time PCR方法，对位于第7号染色体FLNC基因下游53.6 kb处的高同源性假基因（pseFLNC）进行分析。总体而言，50例具有类似细丝蛋白肌病的表型的患者已被筛选FLNC.Results突变：FLNC序列不一致所造成的干扰从pseFLNC被确定和诊断错误，特别是涉及，最常见的致病FLNC p.W2710X突变的检测解决。FLNC和pseFLNC序列之间的错配制表为future use.Conclusions：我们设计了一种策略，允许一个辨别突变发生在功能FLNC从那些窝藏在pseFLNC，从而防止可能的并发症在未来的研究和患者的基因检测。临床化学实验室医学2010;48：1409-14。
Background: Filamin myopathy is a neuromuscular disorder manifesting with predominantly limb-girdle muscle weakness and in many patients with diaphragm paralysis and cardiomyopathy, caused by mutations in the filamin C (FLNC) gene. Molecular diagnosis of filamin myopathy based on direct DNA sequencing of coding exons is compromised by the presence of a high homology pseudogene (pseFLNC) located approximately 53.6 kb downstream of the functional FLNC gene on chromosome 7q.Methods: Molecular cloning, RT-PCR and real-time PCR methods were used to detect sequence differences between the FLNC and pseFLNC that are implicated in known or potential molecular diagnostic errors. Overall, 50 patients with a phenotype resembling filamin myopathy have been screened for mutations in FLNC.Results: FLNC sequence inconsistencies caused by the interference from pseFLNC were identified and diagnostic errors involving, in particular, the detection of the most frequent disease-causing FLNC p.W2710X mutation resolved. Mismatches between the FLNC and pseFLNC sequences were tabulated for future use.Conclusions: We devise a strategy that allows one to discern mutations occurring in the functional FLNC from those harbored in pseFLNC, thus preventing possible complications in future research and patient genetic testing. Clin Chem Lab Med 2010;48:1409-14.