CAJ | 학술논문

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 reduc-tase，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（P<0.01），而纯度与血液样本比较差异无统计学意义（P<0.05）。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。
목적：탐토응용협점막식자채집양본이제취DNA진행고독증보계장애（autism spectrum disorder， ASD）상관기인검측적가행성。방법납입41례ASD환인。채용면식자찰식환인협점막，분-록방-이무순법추제기인조DNA；령채집환인외주정맥혈，용시제합제취기인조DNA。비교량충취재방법획득적기인조DNA총량、농도여순도。응용PCR-한제성매절법대아갑기사경협산환원매（methylenetetra-hydrofolate reduc-tase，MTHFR）기인C677T위점진행기인형분석，비교량충취재법획득적기인분형결과일치성。결과종협점막제취적기인조DNA총량[（5.87±2.58）μg vs.（2.00±0.92）μg]여농도[（143.25±72.78）mg/L vs.（66.68±24.43）mg/L]고우200μL혈액제취적기인조DNA（P<0.01），이순도여혈액양본비교차이무통계학의의（P<0.05）。량충취재법진행MTHFR기인분형결과완전일치，병경Sanger측서험증。결론협점막식자시일충간단、무창、가고적획취기인조DNA방법，가부분체대정맥혈진행ASD상관기인다태성분석。
Objective To investigate the feasibility of buccal mucosa swab method to isolate genomic DNA for au-tism spectrum disorders (ASD)-related genetic screening. Methods Buccal mucosa swabs and blood were collected from 41 children with ASD. Genomic DNA was extracted from either blood by using a commercial genomic DNA kit or buccal mucosa swab by using phenol-chloroform-isoamyl alcohol method. The concentration, total quality and purity of genomic DNA were compared between these two methods. Genotyping of the ASD-related methylenetetra-hydrofolate reductase (MTHFR) gene C677T locus was analyzed using PCR-restriction enzymatic digestion and sanger sequencing was per-formed for validation. Results The total quality [(5.87±2.58)μg vs. (2.00±0.92)μg] and concentration [(143.25±72.78) mg/L vs. (66.68±24.43) mg/L] of genomic DNA extracted from buccal mucosa swab were higher than that form blood (P<0.05), while the purity was not significantly different between these two methods (P>0.05). Genotyping analysis of MTHFR was also consistent between these two methods. Conclusion Buccal mucosa swab is a simple, non-invasive and reliable meth-od to obtain genomic DNA, which can partially replace blood for analysis of ASD-related gene polymorphisms.