CAJ | 학술논문

目的显微操作法联合应用安利快得LV-PCR系统,从而建立一套完整的单细胞DNA分型方法.方法利用显微操作法捕获新鲜口腔上皮细胞,将捕获的细胞直接放置到安利快得玻片反应位点上,加蛋白酶K孵育,然后加入PCR反应混合物进行反应.结果 3个口腔上皮细胞16次均获得完整分型,一个口腔上皮细胞16次中有14次获得完整分型,2次发生部分位点缺失,将该方法应用于两例模拟案例,取得了满意效果.结论联合运用单细胞显微捕获技术和LV-PCR系统对解决微量和混合检材问题具有重要意义.
목적 현미조작법연합응용안리쾌득LV-PCR계통,종이건립일투완정적단세포DNA분형방법.방법 이용현미조작법포획신선구강상피세포,장포획적세포직접방치도안리쾌득파편반응위점상,가단백매K부육,연후가입PCR반응혼합물진행반응.결과 3개구강상피세포16차균획득완정분형,일개구강상피세포16차중유14차획득완정분형,2차발생부분위점결실,장해방법응용우량례모의안례,취득료만의효과.결론 연합운용단세포현미포획기술화LV-PCR계통대해결미량화혼합검재문제구유중요의의.
Objective The micromanipulation method was combined with the AmpliGrid LV-PCR systern to establish a set of single cell DNA genotyping method.Methods Fresh oral epithelium cells were captured by micromanipulation and placed on AmpliGrid reaction position. After incubation with proteinase K,PCR was conducted.Results When three oral epithelium cells were detected,sixteen repeats were all suecessfully finished.For sixteen repeat experiments of single oral epithelium assay,fourteen repeats were completely genotyped,and two repeats were partially genotyped.At last,two simulated cases were successfully detected by this method.Conclusion The combination of single cell micromanipulation with LV-PCR system to resolve low copy number and mixed samples is of great significance