DNA Analysis and Document Examination: The Impact of Each Technique on Respective Analyses

Threatening letters, counterfeit documents, and anonymous notes can commonly be encountered in criminal situations. Such handwritten documents may encourage DNA to transfer from the writer's hands and lower arms when these areas come into contact with the document. As any DNA transferred is likely to be at a low level, sensitive low copy number (LCN) DNA analysis can be employed for testing document exhibits. In this study, we determine locations on the document that are most commonly touched during writing and handling and compare DNA recovery from these sites. We describe the impact of DNA sampling on subsequent document examination techniques including the ESDA^® and likewise the effect of the ESDA^® and two other document examination techniques on subsequent DNA analysis. The findings from this study suggest that DNA results can be obtained through targeted sampling of document evidence, but that care is required when ordering these examination strategies.     

Age Estimation in Living Egyptians Using Signal Joint T‐cell Receptor Excision Circle Rearrangement

Age estimation is one of the challenges in forensic sciences. There are many techniques to estimate the age. Molecular biology approach is one of these techniques. Signal joint T‐cell receptor excision circles gene (sjTRECs), is one of this approach. We aimed to use sjTRECs as a suitable marker for age estimation among Egyptian population. TaqMan qPCR approach was used to quantify sjTREC levels in blood samples obtained from 153 healthy Egyptian individuals ranging from a few weeks to 70 years. Our results showed a significant negative correlation between sjTREC levels and age with p ≤ 0.05. Moreover, the individual's age can be determined through this formula Age = ?30.671+ (?5.998Y) (Y is dCtTBP ? sjTREC) with standard error ±7.35 years. Within the forensic context, sjTREC' levels can be used to estimate the Egyptian individual's age accurately.     

两种DNA提取法对签字笔上脱落细胞STR分型比较

目的比较Chelex-100法和硅珠法两种DNA提取法,在签字笔上附着微量脱落上皮细胞分型中的应用效果。方法 17名志愿者每人使用14支签字笔,每支笔每天使用20min,为期1个月,平均分为两组,分别保存1、3、5、7、14、21和28d,同时运用Chelex-100法和硅珠法两种方法提取签字笔上遗留微量脱落细胞中的DNA,用Identifiler复合扩增系统在AB I 3100遗传分析仪上对这些DNA样品进行STR分型,同时采集上述17名志愿者口腔拭子作为对照。结果以基因座检出个数为指标,使用后签字笔保存1、3、5、7、14、21和28d后,采用Chelex-100法和硅珠法两种方法提取DNA并进行分型检出的基因座个数相比差异均有统计学意义(P0.01);口腔拭子保存1、3、5、7、14、21和28d后,采用Chelex-100法和硅珠法两种方法提取DNA并进行分型检出的基因座个数相比差异均无统计学意义(P0.05)。结论对于微量检材,应用硅珠法提取的DNA分型效果明显优于Chelex-100法,具有较高的应用价值,而在检材量比较多时区别不明显。     

接触DNA在法医实践中的应用研究进展

人体在接触物体后,会在其表面留下少量接触DNA。对接触DNA的检验是目前法医DNA检验技术的热点和难点。接触DNA的检验成功率差异较大,受各环节多重因素影响。影响接触DNA检验成功率的主要因素有接触DNA检材的准确发现、接触DNA的提取与纯化以及扩增策略的选择和结果分析等。本文详述了接触DNA检验过程的进展及研究展望。     

我国法医DNA技术标准化体系建设现状与展望

随着法医DNA技术的飞速发展和广泛应用,客观上必然要求以规范化、标准化的手段对其加以严格管理和正确引导。只有遵循标准化作业,检验所得出的结论才能更科学,更准确,更具有说服力达到为案件提供准确线索和为诉讼提供可信证据的要求。虽然,近年来法医DNA技术标准化工作得到了快速发展,但是仍然存在着不足。本文将对目前法医DNA技术标准的现状、存在问题及展望进行总结。     

罪犯DNA数据库计算机检索系统

目的开发供输入、输出和检索罪犯ＤＮＡ遗传特征的计算机软件包。方法用中文ＶｉｓｕａｌＦｏｘｐｒｏ３０数据库开发软件编程,在建立罪犯ＤＮＡ数据库的基础上,根据ＳＴＲ基因型的类型,设计成显示、增加、编辑、查询、打印和退出等程序模块,并由主程序调用。结果提供一套在中文Ｗｉｎｄｏｗｓ９８环境支持下的、全屏幕菜单提示和操作的罪犯ＤＮＡ数据库计算机检索系统,它能有效地用于罪犯身份的识别     

3种提取胶带粘面汗潜指印中DNA的方法比较

目的比较胶带粘面汗潜指印中DNA提取的方法。方法分别采用硅珠法、QIAMicrokit法、硅珠-QIAMicrokit法提取胶带粘面的汗潜指印中DNA,STR复合扩增,荧光电泳检测。结果用QIAMicrokit法、硅珠-QIAMicrokit法提取胶带粘面汗潜指印中DNA,检测成功率分别为21%和36%。硅珠法检测未获成功。结论硅珠-QIAMicrokit法提取胶带粘面汗潜指印中的DNA比QIAMicrokit法,检验时间更短,检测成功率更高。     

An investigation of two methods of DNA recovery from fired and unfired 9 mm ammunition

Cartridge cases are often recovered from crime scenes involving firearms and, in the United Kingdom (where gun possession is strictly controlled), these are commonly from 9 mm calibre ammunition. The ability to obtain informative DNA profiles from touch DNA on recovered cartridges could have a significant impact on the investigation of that type of offence. However, this avenue may not be routinely considered as investigators in the UK have historically had a low expectation of obtaining useful DNA profiles. This stance may not be unreasonable given that (a) only trace amounts of DNA are likely to have been transferred onto the cartridge cases through handling; and (b) when the cartridge is spent, the potential deterioration of that DNA caused by the act of discharging the weapon.We introduce a novel semi-automatable method using direct lysis for the recovery of DNA from ammunition and compare it with a traditional double-swabbing method (using wet and dry swabs). DNA profiling of the DNA recovered using both methods was carried out using the ESI17 FAST STR system (Promega). This demonstrated a significant increase in DNA recovery using the direct lysis approach, and correspondingly improved STR results.We also investigated the effect on the recovery and profiling of DNA from fired, and unfired, 9 mm cartridges using the direct lysis technique. These results demonstrate that DNA suitable for STR analysis can still be recovered from fired ammunition with only slightly reduced yields compared to unfired ammunition. In these experiments, the handler of the ammunition was most commonly either the sole contributor or the major contributor to the recovered DNA profile.     

DNA isolation success rates from dried and fresh wood samples of selected 20 tropical wood tree species for possible consideration in forensic forestry

The successful isolation of DNA (Deoxyribonucleic Acid) is essential for the investigation process of forestry molecular genetics. Samples used are usually retrieved either from soft or juvenile plant organs because of their excellent DNA source. However, in certain cases, aforesaid samples are hard to obtain, as for forensic purposes. Alternatively, woods possess potential as alternative source of DNA whose extraction method has been developed with varying degrees of success. However, to date, effectiveness on tropical wood grown in Indonesia has not been widely reported. Therefore, objective of this study was to compare the results of DNA isolation of various dried and fresh wood samples by using two isolation methods: Cetyl Trimethyl Ammonium Bromide (CTAB) and Qiagen DNeasy Plant Mini Kit (QDPMK). Extraction results were visualized in agarose gels and quantified using Nanophotometer NP80 Implen which were then amplified using two universal primers: ITS and rbcL for detecting DNA signals. Extraction results from dried wood indicated no visualization in the gel, while fresh wood samples showed thick smeared bands on both extraction methods. Quantity test results denoted higher concentration in CTAB-extracted samples compared to samples extracted using QDPMK, in both types of samples, even though both resulted in optical density ratios outside the range of purity (λ260/280: 1,8–2,0 and λ260/230: 2,0, respectively). Success rates of ITS and rbcL primary amplification in dried wood samples were quite low yet outputs from the two methods did not differ significantly. Meanwhile, outcome of ITS and rbcL amplification on fresh wood samples had a fairly high success.     

Application of fragment analysis based on methylation status mobility difference to identify vaginal secretions

Identifying vaginal secretions attaching or adhering to a suspect’s belongings would be beneficial for reconstructing the events that have taken place during a sexual assault. The present study describes a novel approach to identify vaginal secretions by fragment analysis using capillary electrophoresis, based on the mobility differences of PCR amplicons from bisulfite-treated DNA depending on methylation status. We targeted three genome regions including each of three vaginal secretion-specific methylated CpG sites reported previously: cg25416153, cg09765089, and cg14991487. In all three genome regions, the amplicon peaks for methylated genomic DNA (gDNA) sequences were only detected in vaginal samples, whereas samples of other body fluids (blood, saliva, semen, and deposit on skin surface) only showed amplicon peaks for unmethylated gDNA sequences. In vaginal secretions, the methylation ratio of each of the three targeted regions between samples was variable, while the ratios at the three regions in each sample were similar. Furthermore, commercial vaginal epithelial cells were completely methylated at the three regions. Therefore, vaginal secretion-specific methylation may derive from vaginal epithelial cells present in the sample.In forensic cases with a limited amount of DNA, the reproducibility of a detected peak using the present method is not high due to degradation of DNA by bisulfite treatment and subsequent stochastic PCR bias. However, it was possible to detect peaks from methylated DNA sequences by performing PCR and capillary electrophoresis in triplicate after bisulfite treatment, even when bisulfite treatment was performed using 0.5 ng of gDNA from vaginal secretions. In addition, the level of methylation at each targeted region was found to be stable in vaginal secretions stored for 1 year at room temperature. Therefore, we conclude that detection of the visual peak from vaginal secretion-specific methylated DNA sequence is useful to prove the presence of vaginal secretions. This approach has the potential to analyze multiple marker regions simultaneously, and may provide a new multiplex assay to identify various body fluids.