【摘要】 采用半定量RT-PCR方法检测20例单纯性马蹄内翻足患儿下肢肌肉及肌腱组织中COL1A1基因mRNA的表达,根据COL1A1基因转录调控区-1031bp～+30bp及第1内含子的序列,设计8对引物,PCR扩增后,采用变性梯度凝胶电泳技术筛查突变并测序。半定量RT-PCR结果表明,与正常对照组相比,单纯性马蹄内翻足患儿患侧肌肉及肌腱组织中COL1A1基因表达水平明显上调(t=12.680,P<0.05);经PCR-DGGE筛查并测序发现1名患者存在-161(C→T)的杂合变异,另1名患者存在+274(C→G)的纯合变异。二者均为新发现的变异。提示COL1A1基因转录调控序列变异可能是单纯性马蹄内翻足的致病原因之一。更多还原

【Abstract】 RT-PCR was used to detect the expressions of COL1A1 mRNA in 20 patients with idiopathic congenital talipes equinovarus (ICTEV). The primers were designed by Primer 5 according to sequences of -1 031 bp～ +30 bp and the first intron of COL1A1. PCR-DGGE was used to screen the mutations in COL1A1 gene. Expression of COL1A1 on mRNA lev-els showed significantly higher in patients with ICTEV than in normal persons (t=12.680,P < 0.05). By DNA sequencing,a -161(T→ C) heterozygous mutation and a+ 274(C→G) homozygous mutation were detected,and both were new identified mutations. These results indicated that the mutations in transcription regulator sequences of COL1A1 could cause ICTEV.更多还原