骨骼DNA分析的应用研究

<正> 近年来,各国法庭科学工作者对骨骼DNA检验进行了不断研究与实践,但由于陈旧骨骼受时间和环境因素的影响,对其进行DNA分析仍是目前国际法医界公认的难题。我们前期工作中运用本室改良的硅珠法提取骨组织DNA,对149例骨骼样本中的146例进行STR分型或线粒体测序,并且在此基础上,进一步探讨了骨骼DNA鉴定的难点及其实际应用中应注意的问题,供同仁参考。     

AmpFlSTR MiniFiler™ PCR amplification kit: The new miniSTR multiplex kit

The AmpFlSTR^® MiniFiler™ PCR amplification kit (Applied Biosystems), a new available 8-miniSTR and the sex determining marker Amelogenin multiplex, includes the most common problematic loci (above 200 bp) of the AmpFlSTR^® Identifiler™ PCR amplification kit: FGA, D21S11, D18S51, D13S317, D7S820, D16S539, CSF1PO and D2S1338.Several casework samples with different DNA contents were tested.Results allowed to complete partial Identifiler™ profiles and additional information was achieved in low copy number (LCN) samples, revealing that this miniSTR kit can improve identification of compromised samples.     

全基因组扩增技术及其在法医遗传学中的应用前景

全基因组扩增技术是近年来发展起来的新型PCR技术,它提供了一种从微量基因组DNA获取大量遗传信息的途径,为法医处理微量检材提供了一个有用的工具。本文综述了全基因组技术及其种类和原理和其在法医遗传学方面的应用前景。     

Mini-STRs: A powerful tool to identify genetic profiles in samples with small amounts of DNA

Degraded human remains and crime scene evidences with small amounts of DNA typically reveal incomplete or null genetic profiles when using standard (large) STR amplicons. The technology of mini-STRs, using reduced-size STR amplicons, can help to recover information from these samples. In our Forensic Genetic Service several genetic profiles were obtained or completed using MiniFiler kit (Applied Biosystems) increasing the success rate in sample typing. In all studied cases no inconsistencies were found between profiles obtained with MiniFiler and Identifiler, suggesting that this mini-STR kit can be used to include low copy number (LCN) evidence profiles in STR databases.     

刑事裁判共识引论

共识理论与刑事裁判在问题的起点与终点、解决认识问题的观念、基本要求等方面都有契合之处,表明将共识理论引入刑事裁判问题中是可行的。刑事裁判共识在主体、对象、程度与形成方法上都有其特殊性。传统价值理论的研究主要存在两个缺陷:一是视野局限;二是深度不够。刑事裁判共识理论与传统价值理论具有一致性,可以统一,但又有所超越,所以能够在一定程度上弥补传统价值理论的缺陷。然而,刑事裁判共识理论也与传统价值理论存在冲突,应当根据情况分别对待。     

Genetic analysis of fingerprints—Could WGA or nested-PCR be alternatives to the increase of PCR cycles number?

In this work we aimed to compare the application of increasing PCR cycle number, whole genome amplification and nested-PCR on fingerprints genetic analysis. Results were compared for correct alleles, allele dropin and allelic dropout. We concluded that increasing the number of PCR cycles is yet the best way to attain the required sensitivity.     

Comparison of two whole genome amplification methods for STR genotyping of LCN and degraded DNA samples

The analysis of LCN or highly degraded DNA samples presents a challenge for forensic science. Improving the quantity and/or quality of samples would greatly increase the profiling success rate from LCN and degraded samples. Whole genome amplification (WGA) is one method that has such potential. Two commercially available WGA kits, GenomePlex and GenomiPhi, were investigated for use on LCN and degraded DNA samples. Both kits amplified genomic DNA, producing microgram quantities from sub-nanogram templates. Profiling success of LCN DNA samples was increased, with improvements of over 700% from 10pg template DNA compared to non-WGA-amplified control samples. The amplification success with degraded DNA was also improved by WGA. Degraded DNA was simulated using restriction enzymes to demonstrate that the application of WGA can result in the typing of STR loci that could not previously be amplified. An increase in artefacts, such as stutter alleles and amplification biases, were observed in many samples. Results show that WGA is capable of increasing both the quality and quantity of DNA, and has the potential to improve profiling success from difficult samples in forensic casework.     

MiniFiler~(TM)试剂盒在LCN-STR分型中的应用

目的评估MiniFilerTM试剂盒在LCN-STR分型中的法医学应用价值。方法采用MiniFilerTM与IdentifilerTM试剂盒,对49份常量与39份LCN检材,包括血迹、精斑、骨骼等7类常见检材的检验结果进行比较,并对两种试剂盒的检测灵敏度进行比较。结果在49份常量检材的检验结果中,两种试剂盒的STR分型结果全部一致;在39份LCN生物检材的检验结果中,MiniFilerTM试剂盒获得全部STR分型的有30份,部分分型的5份,阴性的4份;IdentifilerTM试剂盒获得部分STR分型的22份,阴性的17份。MiniFilerTM试剂盒检验成功率明显高于IdentifilerTM试剂盒;MiniFilerTM试剂盒的检测灵敏略高于IdentifilerTM。结论MiniFilerTM试剂盒可显著提高LCN生物检材的检验成功率,适合应用于法庭科学实际检案中LCN生物检材的检验鉴定。     

PCR扩增循环数与低拷贝模板DNA的STR分型

目的探讨PCR扩增循环数对低拷贝模板DNA的STR分型的影响。方法模板DNA(9947A)的不同扩增用量(含低拷贝模板量),采用ProfilerPlus试剂盒,扩增循环数分别为28、30、32、34、38次,3100型基因分析仪(ABI,美国)检测结果。结果循环数从28次增至38次,模板DNA量最低检出量可从0.25ng减少至0.0312ng;先循环28次后,每反应加0.3μlAmpliTaqGoldDNA聚合酶,再循环6次较1次34个循环的检测灵敏度高,相当于1次38个循环的效果。结论增加PCR扩增循环数,可能影响低拷贝模板DNA的STR分型。     

Validation issues around DNA typing of low level DNA

A recent ruling in the Crown Court of Northern Ireland, R v. Hoey, [R v Sean Hoey. 2007, Crown Court of Northern Ireland] has raised questions about the validity of one variant of DNA analysis, often termed LCN. The ruling and subsequent discussion also raises questions about what constitutes validation of a technique.This paper examines what can be achieved in a laboratory based validation study against the Daubert standard and against guidance given in the UK. There is a significant discrepancy between what can be achieved and the Daubert standard but much less of a discrepancy against the UK guidance. Much of the difference relates to differences in word usage, definitional difficulties, and a lack of mutual understanding and communication between the judiciary and forensic scientists. This highlights a gap that needs attention.     

Direct comparison of post-28-cycle PCR purification and modified capillary electrophoresis methods with the 34-cycle “low copy number” (LCN) method for analysis of trace forensic DNA samples

The investigation of samples with low amounts of template DNA remains at the forefront of forensic DNA research and technology as it becomes increasingly important to gain DNA profile information from exceedingly trace levels of DNA. Previous studies have demonstrated that it is possible to obtain short tandem repeat (STR) profiles from <100 pg of template DNA by increasing the number of amplification cycles from 28 to 34, a modification often referred to as “low copy number” or LCN analysis. In this study, we have optimised post-PCR purification techniques applied after only 28 cycles of PCR, as well as using modified capillary electrophoresis injection conditions and have investigated the progressive application of these enhanced approaches. This paper reviews the characteristics of the profiles obtained by these methods compared with those obtained on the same samples after 34-cycle PCR. We observed comparable sensitivity to 34-cycle PCR in terms of the number of profiles with evidence of DNA and the number of allelic peaks per profile and we noted improved peak height and area magnitude with some sample types. Certain parameters reported to be adversely affected in 34-cycle LCN investigations, such as non-donor allele peaks and increased stutter peak ratio, were reduced by this approach. There are a number of advantages for trace samples in progressing from the standard 28-cycle process to the post-PCR processing method as compared to 34-cycle PCR method, including reduced sample consumption, reduced number of PCR amplifications required, and a staged approach to sample processing and profile interpretation.     

Prevalence of mixed DNA profiles in fingernail swabs from autoptic cases

Physical contact between two or more persons can give rise to the transfer of DNA from one person to another and biological material can accumulate under the fingernail hyponichium. The purpose of this study is to value the normal levels of foreign DNA profiles under the fingernail of individuals deceased of a non-violent death, in order to define the usefulness of this approach in the recovery of a suspect's DNA profile. No foreign DNA was found in the majority of the cases.     

宪政视野下的大清新刑律——杨度《论国家主义与家族主义之区别》解读

杨度从宪政的高度看待大清新刑律,使得晚清刑法典论争从法理与礼教的问题之争转向国家与家族的主义之争。在时代的语境中,“个人”通过“国民”这一概念而被国家所吸纳,国家主义乃是看不见独立人格的国民主义。杨度以进化论的观点,将家族视为国家之敌,但对破坏家族是否就能培养出合格的新国民,缺乏充分的论证。他的主义之争加剧了新旧两派的裂痕,使得共识难以达成。     

司法的真相——在法律解释的合法性与妥当性之间

对规范和事实的不同安置,以及由此折射出的对合法性与正义性的不同态度,构成了西方法学史的全部。但至少到目前为止,纠缠于规范与事实、合法与正义之间的争论似乎并未取得令人满意的成果。而从现代西方哲学的商谈理论中我们发现,只有放弃学术立场上的任何一种“独断”,实现规范与事实、法律共同体与相应“听众”之间的合意,并对合法性与正义性进行适合法学语境的重新解读,才能最终缓解甚至消除合法性与正义性(妥当性)之间的冲突。     

Consensus with respect to the fairness of incomes: Differences between social groups

This article deals with consensus on norms on how incomes in a society ought to be distributed. Functionalist theories presume consensus between almost all members of a society. Marxist theories presume at least some dissension, e.g., persons in unfavorable positions will support the achievement principle to a lesser extent and the equality and need principle to a larger measure. In 1987, a vignette study was conducted among a sample of 795 individuals representative of the Dutch population between the ages of 18 and 70. The respondents were presented with a random sample of 15 vignettes and requested to assign a fair income to the described household. A total of 11,394 fair incomes were assigned to the virgnettes. In the analysis the respondents were grouped on value patterns and income position. There was consensus on the main achievement attribute occupational prestige in assigning fair incomes. There was also some dissension discernible between different groups. However, neither of the theories was fully corroborated.     

脱落毛发及毛干DNA的STR分型研究

目的探讨脱落毛发及毛干细胞核DNA提取、含量和STR分型问题。方法对脱落毛发或毛干进行DNA提取和定量,使用低扩增体系、增加循环次数和多次平行扩增等方法扩增DNA样本,采用叠加比较的方法分析STR分型。结果15cm脱落毛发样品DNA含量大于0．3ng的样品占52．8％,STR分型成功率为55．6％;15cm毛干样品DNA含量大于0．3ng的样品占30．6％,STR分型成功率为25％。结论采用增加循环次数、多次平行扩增等LCN—STR分型方法和Mini—STR试剂盒有助于脱落毛发及毛千的STR基因座分型获得。     

PCR扩增3因素对低拷贝模板STR分型的影响研究

目的探讨低拷贝模板(low copy number,LCN)STR扩增方法,提高LCN检材的检验成功率。方法采用Profiler P lusTM试剂盒与9947A对照DNA,改变Taq酶量、体系、循环次数3个因素进行扩增检验,了解各变量对扩增检测的影响。结果对低拷贝模板DNA,单纯增加Taq酶量或反应体系,扩增效率改善不明显;增加循环数,显著提高检验灵敏度;低于0.01ng的模板DNA,同时增加扩增体系、Taq酶量、循环数在一定程度上提高扩增效率。结论对于影响扩增的Taq酶量、体系、循环次数3个因素中,循环数影响最大,但应慎用34次及以上循环数;三者同时增加,对于低于0.01ng模板DNA的扩增可有效改善。     

CAPS理论与人格心理学的整合

米契尔等人在认知-情感人格系统理论中提出了认知-情感单元,认为个体在单元的长期可通达性和单元之间关系的结构上存在差异,这是个体独特性的基础.该理论假设,当个体在不同时间,遇到不同情境时,人格系统会产生具有独特的正面图和情境-行为剖面图.这一理论重新界定了人格系统、状态、特质与动力等概念,在一定程度上调和了人格心理学领域的矛盾,为人格心理学提供了一种整合途径.