法医DNA标准物质备选细胞STR等位基因片段长度的定值

目的 研究建立法医DNA标准物质备选细胞基因组STR基因座等位基因片段长度标准定值的方法.方法 利用有机法提取HPF和HSSM细胞基因组DNA并进行STR复合扩增,将产物进行电泳检测.利用Gene Mapper软件分析电泳结果,记录STR基因座等位基因的数值,并对目前我国公安系统应用较为广泛的DNATyperTM15、IdentifilerTM两种试剂盒扩增产物进行相应的DNA片段长度(bp)统计以及定值.结果 HPF为男性个体细胞,HSSM为女性个体细胞.HPF和HSSM细胞DNATyperTM15系统等位基因片段长度范围分别为126.26±0.05～367.53±0.20bp和125.33±0.07～370.08±0.17bp,IdentifilerTM系统等位基因片段长度范围分别为117.22±0.04～340.02±0.08bp和117.21±0.03～323.86±0.09bp.结论 对STR基因座等位基因片段长度进行标准定值,可为法医DNA标准物质提供有效的溯源途径.     

国产磁珠Wawasye试剂盒在法医DNA中应用

目的探讨国产磁珠Wawasye试剂盒在法医DNA检案中的应用效果。方法利用475份各类常见生物检材测试其效果,并与M48磁珠试剂盒作比较。结果国产磁珠Wawasye试剂盒与M48磁珠试剂盒在检验各类常见生物检材的结果无显著性差异。结论国产磁珠Wawasye试剂盒适用于公安机关DNA实验室。     

Comparison of two methods for isolating DNA from human skeletal remains for STR analysis

Abstract: The quality and efficiency of a standard organic DNA isolation method and a silica‐based method using the QIAGEN Blood Maxi Kit were compared to obtain human DNA and short tandem repeats (STRs) profiles from 39 exhumed bone samples for paternity testing. DNA samples were quantified by real‐time PCR, and STR profiles were obtained using the AmpFlSTR^® Identifiler^® PCR amplification kit. Overall, the silica‐based method recovered less DNA ranging from 0 to 147.7 ng/g (average 7.57 ng/g, median = 1.3 ng/g) than did the organic method ranging from 0 to 605 ng/g (average 44.27 ng/g, median = 5.8 ng/g). Complete profiles (16/16 loci tested) were obtained from 37/39 samples (95%) using the organic method and from 9/39 samples (23%) with the silica‐based method. Compared with a standard organic DNA isolation method, our results indicate that the published silica‐based method does not improve neither the quality nor the quantity of DNA for STR profiling.     

Chelex法和两种磁珠法提取接触DNA效果的比较

目的比较Chelex法、DNA IQ磁珠法、EQ国产磁珠法对接触DNA的提取效果。方法将稀释为10ng、100ng的标准品DNA,分别采用Chelex法、DNA IQ磁珠法、EQ国产磁珠法处理;对30例烟蒂和30例牙刷分别采用Chelex法、DNA IQ磁珠法和EQ国产磁珠法提取DNA,然后进行PCR定量和STR检测。结果Chelex法对DNA的提取无损失,DNA IQ磁珠法、EQ国产磁珠法对DNA的提取均有不同程度的损失;烟蒂、牙刷等检材采用Chelex法提取的接触DNA量和IPC CT值显著高于IQ磁珠法、EQ国产磁珠法,但STR检验成功率却低于IQ磁珠法、EQ国产磁珠法。2种磁珠法提取的DNA量、IPC CT值和STR检验成功率无显著性差异。结论污染轻、杂质少的接触DNA检材,用Chelex法提取最为方便快捷;IQ磁珠法、EQ国产磁珠法更适合污染接触DNA检材的提取及自动化操作。     

法医物证DNA自动化检验技术体系的研究

目的建立自动化工作站同步提取不同种类涉案法医生物检材DNA的新方法。方法选用TECAN Freedom EVO100．4、75—2型自动化提取、加样工作站,采用磁珠法及Chelex-100法对各类涉案生物检材进行DNA提取、PCR扩增、毛细管电泳检测其STR分型,进行比较测试。在“全国公安机关DNA数据库应用系统”中建立并应用实验室信息管理系统（LIMS）模拟实施规范化DNA检案。结果1552份各类检材,采用工作站-磁珠法提取DNA效果最佳,STR检测成功率为95％,工作站-Chelex法为88％;二者分别与其手工提取法比较,成功率无明显差异。92个样本同期检测,自动化工作站较手工操作DNA检案时间可缩减1．25倍。结论工作站域珠法提取涉案检材DNA,可获得满意的STR分型结果。应用LIMS管控,可有效防控污染,明显提高检案效率及鉴定质量。     

纸张上潜在手印三种前处理方法对DNA检验的影响

目的通过比较常见纸张上潜在手印盲提法与显现后精准提取法的接触DNA检出率,探讨常见纸张上接触DNA前处理的优选方案。方法比较五种常见纸张上使用盲提法和显现潜在手印(茚二酮显现法、茚三酮熏显法)后精准提取法采集的接触DNA样本检出率。结果粗糙日历纸盲提的接触DNA检出率为17.8%,通过茚二酮法和茚三酮法显现的潜在手印所提取的DNA检出率分别为75.6%、77.8%;光滑日历纸三种方法所提取的接触DNA检出率为4.4%、11.1%、11.1%;A4复印纸三种方法接触DNA检出率为20%、37.8%、66.7%;牛皮纸三种方法接触DNA检出率为20%、68.9%、64.4%;快递纸袋的三种方法接触DNA检出率为2.2%、6.7%、46.7%。结论不同纸张上潜在手印经显现后接触DNA检出率不同,通过茚二酮或茚三酮显示潜在手印后精准提取DNA的前处理方法相较于盲提法的接触DNA检出率高。实战中可应用此类方法同时获得手印与DNA分型,以有效提高证据力。     

云南大麻DNA的提取及检测初步研究

目的探讨云南大麻DNA的提取及检测方法。方法运用改进的SDS微量法提取大麻总DNA,对所得DNA进行PCR检测。结果用所筛选到的大麻引物检测了云南大麻的DNA遗传标记特征,图谱清晰,结果稳定。结论本方法能有效提取并检测大麻DNA,为实现大麻的品种鉴定、遗传多态性研究及来源追踪提供有价值的科学依据。     

Efficiency of Casework Direct Kit for extraction of touch DNA samples obtained from cars steering wheels

Analysis of STR profiles obtained from touch DNA has been very useful to the elucidation of crimes. Extraction method may be determinant for the recovery of genetic material collected from different surfaces. Vehicle theft is one of the most common crimes in São Paulo city, Brazil, but collection of biological traces in car steering wheels is not considered, because of the belief that profiles generated won’t be able to identify the thief, only the owner. This study aimed to analyze the efficacy of extraction methods for obtaining DNA profiles in samples collected from steering wheels. Eight criminal acts were simulated with 2 different individuals each (mixture of victim and thief), in duplicate, in order to compare two extraction methods: DNA IQ™ and Casework Direct Kit (both Promega Corporation). Genetic material was collected by double swab method and quantified by Quantifiler™Trio (ThermoFisher Scientific). Amplification was conducted with PowerPlex® Fusion System (Promega). It was possible to obtain STR profiles for all experiments. The mixtures were compared with reference profiles to evaluated how many alleles of each donor were observed. Samples extracted with Casework Direct Kit obtained STR profiles with higher averages of alleles for primary and secondary donors (88.7% and 59.9%, respectively) than those extracted with DNA IQ™ (60.4% and 38.1%, respectively). This could be explained by the differences established in the protocols of both methods, since DNA IQ™ is based on successive washes and can result in loss of DNA, whereas Casework Direct Kit minimizes this problem. We concluded that Casework Direct Kit was more efficient for processing touch DNA samples than DNA IQ™.     

TrueAllele® Genotype Identification on DNA Mixtures Containing up to Five Unknown Contributors

Computer methods have been developed for mathematically interpreting mixed and low‐template DNA. The genotype modeling approach computationally separates out the contributors to a mixture, with uncertainty represented through probability. Comparison of inferred genotypes calculates a likelihood ratio (LR), which measures identification information. This study statistically examined the genotype modeling performance of Cybergenetics TrueAllele^® computer system. High‐ and low‐template DNA mixtures of known randomized composition containing 2, 3, 4, and 5 contributors were tested. Sensitivity, specificity, and reproducibility were established through LR quantification in each of these eight groups. Covariance analysis found LR behavior to be relatively invariant to DNA amount or contributor number. Analysis of variance found that consistent solutions were produced, once a sufficient number of contributors were considered. This study demonstrates the reliability of TrueAllele interpretation on complex DNA mixtures of representative casework composition. The results can help predict an information outcome for a DNA mixture analysis.     

The CSI Effect and the impact of DNA evidence on mock jurors and jury deliberations

ABSTRACT

The present research examined the CSI Effect and the impact of DNA evidence on mock jurors and jury deliberations using a 3 (Crime Drama Viewing: low, moderate, high)?×?3 (Evidence: DNA innocent, DNA guilty, no DNA control) design. A sample of 178 jury-eligible college students read a case of breaking and entering. Pre-deliberation, some support for a CSI Effect was found with high viewers’ extent of guilt ratings significantly lower than moderate and low viewers’ in the no DNA control and the DNA innocent conditions. This effect was not present for verdicts. Contrary to a CSI Effect, crime drama viewing was not related to guilt judgments with incriminating DNA evidence. A content analysis of comments made during deliberations found little support for the CSI Effect entering the jury room. Specifically, CSI Effect predictions were not supported when examining the discussion of DNA evidence, expressing DNA opinions, or mentioning missing evidence. Overall, the limited CSI Effect found for individuals was attenuated during deliberation. The alarm raised over a possible CSI Effect influencing jury decision making may be unwarranted.