颊粘膜拭子检测孤独症谱系障碍儿童相关基因多态性的临床应用

发布时间：2018-06-07 11:00

  本文选题：颊粘膜拭子 + 孤独症谱系障碍　； 参考：《中国神经精神疾病杂志》2014年07期

【摘要】：目的探讨应用颊粘膜拭子采集样本以提取DNA进行孤独症谱系障碍(autism spectrum disorder,ASD)相关基因检测的可行性。方法纳入41例ASD患儿。采用棉拭子擦拭患儿颊粘膜,酚-氯仿-异戊醇法抽提基因组DNA;另采集患儿外周静脉血,用试剂盒提取基因组DNA。比较两种取材方法获得的基因组DNA总量、浓度与纯度。应用PCR-限制性酶切法对亚甲基四氢叶酸还原酶(methylenetetra-hydrofolate reductase,MTHFR)基因C677T位点进行基因型分析,比较两种取材法获得的基因分型结果一致性。结果从颊粘膜提取的基因组DNA总量([5.87±2.58)μg vs(.2.00±0.92)μg]与浓度([143.25±72.78)mg/L vs(.66.68±24.43)mg/L]高于200μL血液提取的基因组DNA(P0.01),而纯度与血液样本比较差异无统计学意义(P0.05)。两种取材法进行MTHFR基因分型结果完全一致,并经Sanger测序验证。结论颊粘膜拭子是一种简单、无创、可靠的获取基因组DNA方法,可部分替代静脉血进行ASD相关基因多态性分析。
[Abstract]:Objective to investigate the feasibility of using buccal mucosal swabs to extract DNA for detection of autistic spectrum disordersrelated genes. Methods 41 children with ASD were included. The genomic DNA was extracted from the buccal mucosa by cotton swab and the genomic DNA was extracted by phenol-chloroform-isoamyl alcohol method. The total amount, concentration and purity of genomic DNA obtained by two methods were compared. The C677T locus of methylenetetrahydrofolate reductase (MTHFR) gene was analyzed by PCR- restriction enzyme digestion method. Results the total amount of genomic DNA extracted from buccal mucosa ([5.87 卤2.58) 渭 g vs(.2.00 卤0.92 渭 g] and the concentration ([143.25 卤72.78)mg/L vs(.66.68 卤24.43)mg/L]) were higher than that of 200 渭 L blood samples. The results of MTHFR genotyping by two methods were identical and confirmed by Sanger sequencing. Conclusion buccal mucosal swab is a simple, noninvasive and reliable method for obtaining genomic DNA, which can partly replace venous blood for ASD related gene polymorphism analysis.
【作者单位】： 山东大学齐鲁儿童医院儿童研究所;山东大学齐鲁儿童医院 山东省孤独症儿童康复中心;
【分类号】：R749.94
，

本文编号：1990940