国际遗传学杂志
國際遺傳學雜誌
국제유전학잡지
INTERNATIONAL JOURNAL OF GENETICS
2008年
3期
180-182
,共3页
陈玲%刘超%王慧君%邱平明%杨电
陳玲%劉超%王慧君%邱平明%楊電
진령%류초%왕혜군%구평명%양전
多重置换扩增反应%改进型扩增前引物延伸反应%全基因组扩增%STR分型
多重置換擴增反應%改進型擴增前引物延伸反應%全基因組擴增%STR分型
다중치환확증반응%개진형확증전인물연신반응%전기인조확증%STR분형
Multiple displacement amplification%Improved primer extension preamplification%Whole genome amplification%STR genotyping
目的 应用多重置换扩增(MDA)技术和改进型扩增前引物延伸(IPEP)技术对法医学微量DNA进行全基因组扩增,比较两种方法的STR分型效果及法医学应用价值.方法 用MDA、IPEP方法分别对不同模板量DNA进行扩增,扩增产物用实时荧光定量PCR技术定量、用AmpFLSTR Identifiler(R)试剂盒检测基因型.结果 MDA方法可对模板DNA增加103～106倍,IPEP方法可增加25～310倍.为获得完整准确的分型结果,MDA最低需1 ng基因组DNA,IPEP最低需0.05 ng基因组DNA.当基因组DNA为0.01 ng～0.1 ng时,IPEP产物、MDA产物的平均基因座检出数均高于未经全基因组扩增的DNA,其中IPEP产物的平均基因座检出数高于MDA产物.结论 MDA方法、IPEP方法均可提高微量检材的STR分型效果.MDA方法的产量高于IPEP方法;IPEP方法的灵敏度高于MDA方法,且对微量DNA的STR分型效果优于MDA方法,因此更适于法医学痕量DNA检测.
目的 應用多重置換擴增(MDA)技術和改進型擴增前引物延伸(IPEP)技術對法醫學微量DNA進行全基因組擴增,比較兩種方法的STR分型效果及法醫學應用價值.方法 用MDA、IPEP方法分彆對不同模闆量DNA進行擴增,擴增產物用實時熒光定量PCR技術定量、用AmpFLSTR Identifiler(R)試劑盒檢測基因型.結果 MDA方法可對模闆DNA增加103～106倍,IPEP方法可增加25～310倍.為穫得完整準確的分型結果,MDA最低需1 ng基因組DNA,IPEP最低需0.05 ng基因組DNA.噹基因組DNA為0.01 ng～0.1 ng時,IPEP產物、MDA產物的平均基因座檢齣數均高于未經全基因組擴增的DNA,其中IPEP產物的平均基因座檢齣數高于MDA產物.結論 MDA方法、IPEP方法均可提高微量檢材的STR分型效果.MDA方法的產量高于IPEP方法;IPEP方法的靈敏度高于MDA方法,且對微量DNA的STR分型效果優于MDA方法,因此更適于法醫學痕量DNA檢測.
목적 응용다중치환확증(MDA)기술화개진형확증전인물연신(IPEP)기술대법의학미량DNA진행전기인조확증,비교량충방법적STR분형효과급법의학응용개치.방법 용MDA、IPEP방법분별대불동모판량DNA진행확증,확증산물용실시형광정량PCR기술정량、용AmpFLSTR Identifiler(R)시제합검측기인형.결과 MDA방법가대모판DNA증가103～106배,IPEP방법가증가25～310배.위획득완정준학적분형결과,MDA최저수1 ng기인조DNA,IPEP최저수0.05 ng기인조DNA.당기인조DNA위0.01 ng～0.1 ng시,IPEP산물、MDA산물적평균기인좌검출수균고우미경전기인조확증적DNA,기중IPEP산물적평균기인좌검출수고우MDA산물.결론 MDA방법、IPEP방법균가제고미량검재적STR분형효과.MDA방법적산량고우IPEP방법;IPEP방법적령민도고우MDA방법,차대미량DNA적STR분형효과우우MDA방법,인차경괄우법의학흔량DNA검측.
Objective To apply multiple displacement amplification (MDA)to improve primer extension preamplification (IPEP) for whole genome amplification (WGA) and to compare their effects on forensic DNA analysis. Methods DNA samples of varying amounts were for WGA based on MDA and IPEP.WGA products yield was evaluated by real-time quantitative PCR and STR genotyping performance was determined with AmpFLSTR? Identifiler(R) Kit. Results The DNA quantity was increased about 103～106 folds by MDA and 25～310 folds by IPEP. The least genome DNA amount is 1ng for MDA and 0.05ng for IPEP to obtain accurate genotypes of all loci. IPEP products and MDA products of 0.01 ng～0.1 ng DNA exhibited more loci observed than original DNA that was not for WGA,and IPEP products exhibited more loci observed than MDA products. Conclusion Both MDA and IPEP can improve STR genotyping of forensic minute DNA. The yield of MDA is higher than IPEP but the sensitivity of IPEP is higher than MDA and the STR genotyping effect of IPEP products is better than MDA products so IPEP is better for forensic trace DNA analysis.
