插入缺失位点和miniSTR在甲醛固定石蜡包埋组织中的应用检测

Application detection of insertion/deletion locus and miniSTR in formalin fixed paraffin embedded tissue

目的:研究插入缺失位点InDel与mini短串联重复序列(STR)遗传标记对甲醛固定石蜡包埋组织应用的价值。方法:19份甲醛固定石蜡包埋组织作为疑难检材,应用3种常染色体遗传标记多重扩增系统,即InDel系统(Investigatior■DIPplex Kit)、miniSTR系统(华夏白金扩增试剂盒)和STR系统(Goldeneye®20A),对STR分型不完整或失败的检材,再分别用InDel和miniSTR系统进行DNA分型检测,比较两者的分型质量和检出率。结果:19份甲醛固定石蜡包埋组织经Goldeneye 20A试剂盒检测,均分型不完整。应用miniSTR系统检测显示,7份样本的检出率100%,其中,D19S433基因座的检出率为100%,D5S818、D13S817和D1S1656基因座的检出率最小,为36.84%。应用InDel系统检测发现,1份样本InDel位点电泳分型完整,所有样本的位点丢失率均小于50%,其中7份样本的位点丢失率低至10%以下。结论:应用遗传标记对甲醛固定石蜡包埋组织这类疑难降解检材进行检测,插入缺失多态性位点InDel与miniSTR可以作为联合检测标记。

Objective:This paper studies the application value of Insertion/Deletion(InDel)and miniSTR genetic marker in difficult degraded sample,such as formalin fixed paraffin and embedded tissue,so as to guide the selection of genetic marker in practical application.Methods:A total of 19 formalin fixed paraffin and embedded tissue were genotyped with three systems:the InDel system(Investigatior■DIPplex Kit),miniSTR system(Huaxia platinum amplification Kit),and STR systems(Goldeneye 20 A).For the incomplete or failed STR,DNA typing were performed by InDel and miniSTR systems to compare their ty ping quality and detection rate.Results:Nineteen formalin fixed and paraffin embedded tissues were detected incomplete by the Goldeneye 20A kit.The miniSTR system was applied,and the detection rate of seven samples was 100%,among which D19S433 locus were 100%,and D5S818,D13S817,and D1S1656 locus were the smallest,36.84%.The DNA typing of one sample was complete with the InDel system.The loss rate of all samples were less than 50%,and that of seven samples were less than 10%.Conclusion:InDel and miniSTR can be used as the combined detection markers for the detection of formalin fixed and paraffin embedded tissues that are hard with degradation.