棉织物上血斑洗涤后无损取材方式及DNA提取方法研究  

Study on nondestructive sampling and DNA extraction method of laundered bloodstains on cotton fabricss

作　　者：李泽琴 刘娜 原芳 张迪 王刚 贠克明[1] 严江伟 张更谦[1] Li Zeqin;Liu Na;Yuan Fang;Zhang Di;Wang Gang;Yun Keming;Yan Jiangwei;Zhang Gengqian(School of Forensic Medicine,Shanxi Medical University,Jinzhong 030600,China;Beijing Institute of Information Technology,Beijing 100094,China)

机构地区：[1]山西医科大学法医学院,山西晋中030600 [2]北京市信息技术研究所,北京100094

出　　处：《中国法医学杂志》2023年第3期284-289,共6页Chinese Journal of Forensic Medicine

基　　金：山西省自然科学研究面上项目(20210302123313)。

摘　　要：目的探究不同无损取材方式及DNA提取方法对棉织物上洗涤血斑DNA分析成功率的影响。方法采用棉签擦拭法、植绒拭子擦拭法、脱落细胞粘取器粘取法富集洗涤棉织物上的血痕。分别采用Chelex 100法、过柱法和磁珠法提取DNA。采用常规PCR和复合PCR技术扩增目标片段并分别运用琼脂糖凝胶电泳和毛细管电泳技术检测目标产物。结果植绒拭子擦拭法取材的效果最差,通过脱落细胞粘取器粘取法获得的DNA浓度、STR分型成功率均高于棉签擦拭法,且粘取法操作较简单,实践中优先推荐选择粘取法作为该类检材的取材方式。此外,DNA浓度和STR分型结果均表明过柱法提取DNA的效果优于Chelex法,而磁珠法在3种DNA提取方法中效果最差,建议实践中选择过柱法作为此类检材DNA提取的首选方法。结论针对棉织物检材上洗涤血痕无损取材及DNA分析,优先推荐选择脱落细胞粘取器粘取法和过柱法分别进行样本取材以及DNA提取。Objective To explore the influence of different non-destructive sampling methods and DNA extraction methods on the success rate of DNA analysis of laundered bloodstains deposited on cotton fabrics.Methods Blood trace deposited on the washed cotton fabrics was obtained using cotton swab,flocked swab,or exfoliated cell adhesion method.DNA was extracted by the methods of Chelex 100,spin column or magnetic bead.Target fragments were amplified by conventional PCR and multiplex PCR technology and the products were separated by agarose gel electrophoresis and capillary electrophoresis,respectively.Results The flocked swab method showed the worst sampling effect.The DNA concentration and the success rate of STR typing obtained through adhesion method were higher than those obtained by cotton swab method,and the operation of adhesion method was simpler.Therefore,the adhesion method is recommended as the non-destructive sampling method in practice.In addition,the results of DNA concentration and STR typing showed that the DNA extraction efficiency of the spin column method was better than that of the Chelex method,while the magnetic bead method was the least effective among the three methods.It is suggested to choose spin column method as the first choice for DNA extraction of such samples in practice.Conclusion For the non-destructive analysis of the laundered bloodstains deposited on cotton fabrics,the exfoliated cell adhesive tool and the spin column method are recommended for sampling and DNA extraction,respectively.

关 键 词：法医检验 洗涤血痕 无损取材 DNA提取 DNA分析 

分 类 号：D919.2[医药卫生—法医学]