毛干检材STR基因分型成功率的影响因素研究  

Study on influencing factors of success rate of STR genotyping from hair shafts inspection material

作　　者：郑阳阳 郑文彦 吴伟斌 杜蔚安 ZHENG Yangyang;ZHENG Wenyan;WU Weibin;DU Weian(Guangdong Huamei Zhongyuan Biotechbology Co.,Ltd.,Foshan,Guangdong 528000,China)

机构地区：[1]广东华美众源生物科技有限公司,广东佛山528000

出　　处：《现代医药卫生》2021年第6期950-953,共4页Journal of Modern Medicine & Health

摘　　要：目的分析自然脱落毛干检材短串联重复序列(STR)分型成功率的影响因素。方法2019年10月收集15名健康志愿者毛干样本,采用chelex-100法、磁珠法提取纯化DNA,并进行Qubit荧光系统定量测定,利用自设引物及AGCU Expressmarker 16CS STR分型试剂盒进行扩增、分型。结果从15名健康志愿者选取的8名健康志愿者毛干DNA含量为0.01~0.78ng,利用毛干DNA为模板的大小片段扩增结果具有明显差异,证明其存在降解;黑色素对聚合酶链反应具有抑制作用,抑制质量浓度为36ng/μL。结论毛干STR分型的成功率与DNA含量低、DNA降解及黑色素抑制有关。通过降低提取过程中DNA损失率、采用短扩增子基因分型试剂或优化聚合酶链反应体系以缓解黑色素抑制等途径有助于改善毛干分型结果。Objective To analyze the influencing factors of the success rate of STR typing from natural abscisic hair shafts inspection material.Methods The hair shafts samples of 15 healthy volunteers were collected in October 2019.The chelex-100 method and magnetic bead method were adopted to extract and purify DNA.The Qubit fluorescent system quantitative determination was performed.The self-designed primer and the AGCU Expressmarker 16 CS typing kit were used to conduct the amplification and genotyping.Results The DNA content of hair shafts in 8 healthy volunteers selected from 15 healthy volunteers was 0.01-0.78 ng.The large and small fragment amplification results by using the hair shafts DNA as the template had significant difference,which confirmed that there existed the degradation.Melanin had the inhibiting effect on PCR,and the inhibiting mass concentration was 36 ng/μL.Conclusion The success rate of STR genotyping from hair shafts is related with the low DNA content,DNA degradation and melanin inhibition.The approaches such as decreasing the DNA loss rate during the extraction process,adopting the short amplicon genotyping reagent or optimizing PCR system for relieving the melanin inhibition is conducive to improve the hair shafts typing results.

关 键 词：毛发 法医学 司法鉴定 基因型 短串联重复序列 

分 类 号：R89[医药卫生—法医学]