CAJ | 학술논문

目的：建立并优化用于血浆microRNA（ miRNA）检测分析的miRNA快速提取方法。方法使用含表面活性剂的提取液和加热方法提取血浆miRNA，采用RNA酶抑制剂抑制RNA降解，利用检测miRNA的poly（A）加尾、逆转录反应和实时定量PCR法检测血浆miRNA表达水平，评估方法的可行性。结果成功建立了提取液和加热提取miRNA一步法，实时定量PCR结果表明该方法具有良好的扩增特异性、可重复性，方法稳定可靠。结论提取液和加热一步法简便、快速、低成本，血浆用量少，为临床血浆miRNA检测分析提供了一种新的技术方法。
목적：건립병우화용우혈장microRNA（ miRNA）검측분석적miRNA쾌속제취방법。방법사용함표면활성제적제취액화가열방법제취혈장miRNA，채용RNA매억제제억제RNA강해，이용검측miRNA적poly（A）가미、역전록반응화실시정량PCR법검측혈장miRNA표체수평，평고방법적가행성。결과성공건립료제취액화가열제취miRNA일보법，실시정량PCR결과표명해방법구유량호적확증특이성、가중복성，방법은정가고。결론제취액화가열일보법간편、쾌속、저성본，혈장용량소，위림상혈장miRNA검측분석제공료일충신적기술방법。
Objective To develop and optimize a new method to extract miRNAs from plasma.Methods miRNAs were extracted from plasma by mixing it with the extraction solution that contained surfactant and by heating .Then the ribonuclease inhibitor was added into the extraction to prevent RNAs from degradation .The expression level of each miRNA was detected by real-time quantitative PCR in oder to evaluate the feasibility of this method .Results A method which extracted miRNAs from plamsa in just one step was established .The specificity , reproducibility and stability of this method have been demonstrated by real-time quantitative PCR .Conclusion The one-step method is simple , inexpensive , and plasma-saving.It seems like a new method for clinical examination of miRNAs from plasma .