硅珠法提取微量检材DNA质量的研究-现代医学

硅珠法提取微量检材DNA质量的研究

单位：南京红十字血液中心体检采血科, 江苏南京 210003

分类号：R446.6

出版年·卷·期（页码）：2024·52·第六期（920-925）

摘要：

目的： 运用Goldeneye 20A 试剂盒评估硅珠法提取微量检材DNA的质量。方法： 运用硅珠法对40名健康志愿者提供的血液、口香糖、烟蒂、指甲、牙刷样本提取DNA，Goldeneye 20A试剂盒扩增后分析提取DNA的质量。结果： 与志愿者提供的血样比对，其他微量样本经硅珠法提取后获得的DNA，在检材适应性方面采集成功率(92.50%)最低的牙刷样本经检验与血液样本扩增成功率相比差异无统计学意义(P=0.241)；1 μL血液样本即使稀释至50倍以内再采用硅珠法提取DNA，也可以获得足够的DNA用于短串联重复序列(short tandem repeat,STR)分型，与9947A标准品阳性对照基因座(20个)和等位基因数(40个)比较分型结果正确；4种微量检材经硅珠法提取的DNA扩增电泳后，20个基因座图谱峰值均可超过200 RUF，且重复检验图谱峰值水平相当，分型结果同血液样本一致。硅珠法提取DNA能满足片段多态性分析的需求，并且均获得较好的STR分型图。结论： 硅珠法可以运用于多种微量检材的DNA提取，有较广泛的适应性，能获得满足于STR检测要求的DNA。

Objective: To evaluate the quality of DNA extracted by silica using Goldeneye 20A Kit. Methods: DNA was extracted from blood, chewing gum, cigarette butts, nails and toothbrush samples provided by 40 healthy volunteers by silicon beads method, and the quality of extracted DNA was analyzed after amplification using Goldeneye 20A kit. Results: Compared with the blood sample provided by the volunteers, collection success rate of DNA obtained from other trace of samples by silica bead extraction was 92.50%. The toothbrush sample with the lowest sample adaptability was not significant compared with blood sample in terms of the success rate(P=0.241); even one microliter of blood sample was diluted to 50 times before the DNA was extracted by the silicon bead method, we still could obtain sufficient DNA for STR typing, typing results of 20 control locus and the 40 alleles were correct compared with the standard 9947A positive; after electrophoresis of DNA extracted by silica bead method, the spectrum of 20 locus peaks exceeded 200 RUF, and repeated test achieved comparable peak levels. The subtyping results were consistent with the blood samples. DNA extraction by silicon bead method can meet the requirements of fragment polymorphism analysis, and obtain good short tandem repeat(short tandem repeat, STR) typing plots. Conclusion: The silicon bead method can be used in the DNA extraction of a variety of microsamples, which has a wide range of adaptability, and can obtain the DNA that meets the requirements of STR detection.

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