PrepFiler Express BTA~(TM)裂解液联合硅珠法快速提取骨骼DNA

目的建立方便、快捷的提取骨骼DNA的方法。方法对15份长骨样本清洗消毒后,采用电钻钻取骨屑,使用PrepFiler Express BTA~(TM)裂解液进行裂解消化后,应用手工硅珠吸附纯化进行DNA提取检验。结果有14份样本获得满意的STR分型,仅1份样本未获得STR分型,检出率为93.33%。结论本研究建立的骨骼DNA快速提取方法操作简单、便捷、有效,适应性强,值得推广应用。     

Application of RapidHITTM 200 System in Forensic MedicineCSCD

Objective: To validate the analysis capability of RapidHITTM 200 system for four kinds of routine forensic samples and the recyclable capability of template, template DNA and PCR products in the process of twice duplicate detection. Methods: The buccal swabs underwent the test twice by RapidHITTM 200 system, and the template DNA and PCR products that arose in the system were also tested for two times. After four kinds of routine forensic samples were detected by RapidHITTM 200 system, the follow- up tests of the template, template DNA and PCR products that arose in the system were performed. Results: The STR loci could be detected in the buccal swabs by the system for the first time. However, part of the STR loci lost during the second test. And the peak value obtained in the second test was significantly reduced than the one in the first time. The average STR loci detection rates of the template DNA and PCR products were both less than 50% in the second test, which were significantly reduced than that in the first test. In addition, the analysis capability of the system for the tissues and buccal swabs was better than that for the blood and cigarette butts. Compared with the first test, the STR loci detection rate of the tested items, template DNA and PCR products decreased with the numbers of tests. Conclusion: RapidHITTM 200 system is more effective in retesting buccal swabs than other samples, whereas the items, DNA template, PCR products obtained in the first and second time cannot be directly used for the further application and study of forensic medicine. © 2018 by the Editorial Department of Journal of Forensic Medicine.     

改良差异裂解法结合硅珠法提取混合斑精子DNA

目的采用改良差异裂解法结合硅珠法提取混合斑中精子DNA,并评价其应用价值。方法收集52例经常规差异裂解法检验含有女性分型的混合斑检材,采用改良差异裂解法结合硅珠法提取精子细胞DNA,IdentifilerTM试剂盒进行PCR扩增检验。并将常规差异裂解法结果作为对照。结果52例混合斑检材中,采用改良差异裂解法结合硅珠法检出单一男性精子成分有38例,男性分型检出率达到98．08％。结论改良差异裂解法结合硅珠法适合提取混合斑中精子DNA。     

Evaluation of five DNA extraction systems for recovery of DNA from bone

Five DNA extraction systems were assessed for their DNA extraction efficiency on samples of fresh pig bone. Four commercially available silica-based extraction kits (ChargeSwitch^® gDNA Plant Kit (Life Technologies), DNA IQ^TM System Kit (Promega), DNeasy^® Blood & Tissue Kit (Qiagen) and PrepFiler^® BTA Forensic DNA Extraction Kit (Life Technologies)) and a conventional phenol-chloroform method were tested in this study. Extracted DNA samples were quantitated with GoTaq^® qPCR Master Mix (Promega) using an Applied Biosystems^® 7500 Real-Time PCR System and the extracts were amplified using an in-house multiplex system. The phenol-chloroform extraction produced higher yields of DNA than the silica-based extraction methods. Among the silica-based extractions ChargeSwitch^® gDNA Plant Kit recovered the highest amounts of DNA. However, all methods produced DNA that could be amplified and none of the extracts contained any detectable inhibition.