CAJ | 학술논문

目的对常规TRIzol法提取血清中microRNA(miRNA)的方法进行改进,以提高血清miRNA的提取效率.方法运用常规TRIzol法和改进方法,分别提取健康人血清中的miRNA.改进条件为分离血清后再分别进行超声、10％ SDS(十二烷基硫酸钠)或者10％ SDS联合超声进行预处理.以实时荧光定量PCR检测血清中miRNA-218和miRNA-346的含量,通过比较miRNA-218和miRNA-346的相对表达量来鉴定不同方法对miRNA检测值的影响,从而反映miRNA的检出效率.结果超声组与常规法组的检出效率比较差异无统计学意义(P＞0.05);SDS组与SDS联合超声组检出效率均优于常规组(P＜0.05),此两组改进方法可使荧光定量PCR检测中miRNA-218和miR-NA-346的相对表达量明显提高.而SDS联合超声组检出效率比单纯SDS组更好(P＜0.05).结论 10％SDS联合超声预处理后,可有效提高目的血清miRNA的检出效率.
목적 대상규TRIzol법제취혈청중microRNA(miRNA)적방법진행개진,이제고혈청miRNA적제취효솔.방법 운용상규TRIzol법화개진방법,분별제취건강인혈청중적miRNA.개진조건위분리혈청후재분별진행초성、10％ SDS(십이완기류산납)혹자10％ SDS연합초성진행예처리.이실시형광정량PCR검측혈청중miRNA-218화miRNA-346적함량,통과비교miRNA-218화miRNA-346적상대표체량래감정불동방법대miRNA검측치적영향,종이반영miRNA적검출효솔.결과 초성조여상규법조적검출효솔비교차이무통계학의의(P＞0.05);SDS조여SDS연합초성조검출효솔균우우상규조(P＜0.05),차량조개진방법가사형광정량PCR검측중miRNA-218화miR-NA-346적상대표체량명현제고.이SDS연합초성조검출효솔비단순SDS조경호(P＜0.05).결론 10％SDS연합초성예처리후,가유효제고목적혈청miRNA적검출효솔.
Objective To investigate which methods can promote the serum microRNA extract production.Methods The sera of healthy persons were treated with 10％ SDS (sodium dodecyl sulfate) and/or sonication at first,then extracted miRNA by routine Trizol method,or the serum miRNA was just extracted by routine Trizol method.The contents of serum miRNA-218 and miRNA-346 were detected by quantitative reverse transcription polymerase chain reaction (qRT-PCR) technique,and qRT-PCR relative quantities among different groups were compared to evaluate influence of different methods on microRNA extract efficacy.Results Compared to routine Trizol extraction method,the qRT-PCR relative quantities of miRNA-218 and miRNA-346 were improved when serum was treated with 10％ SDS and/or sonication at first (P ＜0.05),which was most obviously improved when treated with 10％ SDS combined with sonieation.Conclusions The method that serum was treated with SDS incubation and sonication at first can promote sensitivity of serum microRNA detection.