单核苷酸多态性与法医学DNA分型技术

单核苷酸多态性(single nucleotide polymorphism,SNP)分型技术越来越成为法医学领域关注的热点,它在研究Y染色体或线粒体单倍型以及DNA表型的分析中具有重要应用价值。本文着重比较分析了SNP技术与片段长度多态性技术之间的优劣,同时就当前STR位点识别概率与所需选择的SNP位点数进行探讨。此外,本文还就各类SNP分型方法的优缺点及其法医学应用进行了论述。     

法庭科学DNASTR分型标准物质初探

本文介绍了标准物质及法医DNA标准物质的概念,并就目前法庭科学DNASTR分型技术所需要的标准物质的制备及应用做一概述,以对法医DNA标准物质的生产提供参考。     

Basic issues in forensic DNA typing

DNA analysis has become the standard method in forensic stain typing (termed DNA profiling). In contrast to conventional serological methods, any human tissue or body fluid can be analysed by DNA profiling as long as it contains nucleated cells. The majority of genetic systems studied at the DNA level are derived from “non-coding” portions from the human genome, and are located either in the vicinity of expressed (coding) genes or in stretches of DNA sequences interspersing with the genes. The typing results are usually recorded as DNA fragment lengths or “alleles” indicating the number of core repeat elements for short tandem repeat systems. These typing results do not contain any useful information which might reveal genetic traits or predispositions for inherited disease about the individual studied. Typing systems for DNA profiling are predominantly selected according to criteria related to the robustness for typing of (potentially degraded) forensic specimens, the degree of genetic polymorphism (which influences the chance to exclude a wrongfully accused person), and the amenability to standardisation as a basis to obtain reproducible results.     

HLA-A位点DNA分型及其法医学应用

应用聚合酶链反应0寡核苷酸探针（PCR－SSO）斑点杂交技术，对222名辽宁地区汉族人群无关个体进行HLA－A基因检测，研究中国辽宁地区汉族群体的HLA－A座位基因分布状况。共检出HLA－A等位基因24个，其中以等位基因HLA－A0201最为常见，频率为0．2635；依次是2402101和1101，等位基因频率分别为0．1847和0．1262.理论杂合度为87％，个人鉴别机率为92％，非父排除率为73．3％。在中国辽宁汉族中检出73种基因型，对观察值和期望值进行X2检验，符合Hardy－Weinberg平衡定律（x2＝6．28，df＝9，0．5＜P＜0．75）。家系分析结果表明按照孟德尔方式遗传。提出的中国辽宁汉族HLA－A等位基因的遗传基因情况，可用于法医学个人识别和亲子鉴定。人类学，HLA相关疾病，及器官移植研究。     

DNA分型实验室管理应重视的几个问题

1985年Gill等第一次报导将DNA指纹图用于个体讽别并获得成功后，DNA分型技术在个体识别和亲子鉴定中的应用潜力开始为法医界所注目【'］．随着该项技术的日益完善，它极大地提高了生物学检材（血液，血痕，精斑，混合斑，毛发和指甲等）在法医物证学鉴定过程中的价值，并广泛应用于法医鉴定实践中．同时，法医界又面临着这样一个问题：如何对有关的DNA分型实验室进行管理监督，以保证其能更好地为司法实践服务．本文从DNA分型实验室人员设置和要求，DNA分型技术标准化以及与传统血型血清学的关系等方面并结合国外进展作如下探讨．ID…     

47个SNPs复合检测方法的建立及其法医学应用

目的建立47-plexSNPs复合检测方法，评价其在法医学中的应用价值。方法筛选46个常染色体SNPs和1个Y—SNPs，使用2个检测体系分别对47个SNPs进行单管内复合PCR扩增，采用荧光标记单碱基延伸法和毛细管电泳检测技术进行分型检测；并用建立的方法对260份广东地区无关个体血样进行47个SNPs分型。结果建立的47-plex SNPs的复合检测体系灵敏度高，种属特异性好；260名个体所有SNPs均能准确分型，群体内基因型频率分布均符合Hardy—Weinberg平衡，累积个人识别率大于0．9999，累积非父排除率为0．99982，累积偶合率为6．24×10一。结论本文47-plex SNPs复合检测方法能同时对47个SNPs进行快速、准确的检测，在法医学个体识别鉴定中具有良好的应用前景。     

DNA双股螺旋结构与迅猛发展的法庭科学——纪念人类DNA双股螺旋结构发现50周年

自1953年Watson和Crick发表DNA双股螺旋结构,它为人类生命科学,尤其是法庭科学的发展带来了历史性的变化。法庭科学从个体识别和亲子鉴定的排除到认定,经历了DNA指纹图、AMP-FLP及SNP的3个阶段。同时线粒体DNA的应用也在广泛开展。另外数据库的建设和完善成为法庭科学的发展方向。总之,DNA双股螺旋结构对法庭科学革命性变化起了决定性的作用。     

The collaborative study on typing group-specific component by eight forensic science laboratories

This report describes a collaborative study on typing group-specific component (GC), conducted between the Central Research and Support Establishment and the forensic science laboratories of England, Wales and Northern Ireland. A population study (n=114) was performed. Fifty blood donors were selected to provide a distribution, slightly biased from normal, in favour of the GC 1F-1F and GC 1F-1S phenotypes. A protocol was devised for preparing large bloodstains. The strongest GC bands were obtained from the edge of a stain after the blood had been treated with K+/EDTA. Each laboratory received a representative portion of the large bloodstains for GC typing. Five of the eight laboratories correctly grouped all the bloodstains. No errors directly attitributable to the system were recorded in over 800 tests, indicating that GC in bloodstains can be typed reliably using the combination of isoelectric focusing in ultrathin narrow pH interval gels followed by immunofixation and silver staining.     

DNA分析技术在法科学中的应用及展望

本综述对DNA指纹、PCR -VNTR、PCR -STR、PCR -mtDNA测序等技术的发展 ,以及其在法科学中的应用领域和发展前景作了系统的阐述。认为由于DNA分析技术所具有的特点 ,使之已成为现今法科学生物检材检验的主要手段之一。阐述了现阶段DNA分析技术已向标准化、自动化和高鉴别机率方向发展 ,以及建立DNA罪犯数据库的必要性和应用价值     

DNA分析技术在法科学中的应用及展望

本综述对ＤＮＡ指纹、ＰＣＲ－ＶＮＴＲ、ＰＣＲ－ＳＴＲ、ＰＣＲ－ｍｔＤＮＡ测序等技术的发展,以及其在法科学中的应用领域和发展前景作了系统的阐述。认为由于ＤＮＡ分析技术所具有特点,使之已成为责令法科学生物检材检验的主要手段之一。阐述了现阶段ＤＮＡ分析技术已向标准化、自动化和高鉴别机率方向发展,以及建立ＤＮＡ罪犯数据库的必要性和应用价值。     

Cell line DNA typing in forensic genetics--the necessity of reliable standards

The incorporation of reference DNA is crucial to the validation of any DNA typing protocol. This paper aims to provide a panel of reference DNAs for actual forensic profiling strategies, i.e. autosomal and gonosomal STR typing as well as mtDNA sequencing. We have characterised three human lymphoid cell lines, GM9947, GM9948 and GM3657, and considered 58 autosomal and gonosomal microsatellites as well as the mitochondrial control region sequence. Well-established markers and STRs recently developed for forensic use were involved. K562 DNA samples which we purchased from two different suppliers were also analysed. They revealed conflicting results with regard to the ChrX STR marker genotype. Hence, we suggest that K562 is no longer used for the calibration of profiling techniques. Our investigation establishes a panel of one female and two male DNA samples as an STR allelic ladder calibration tool and offers information on six alleles of each autosome (AS) marker, three alleles of each X chromosome (ChrX) marker and two alleles of each ChrY marker. In addition, sequences of the mitochondrial control region of the three DNAs are communicated in order to provide sequencing quality control.     

Expanding forensic science through forensic intelligence

Research and Development (‘R&D’) in forensic science currently focuses on innovative technologies improving the efficiency of existing forensic processes, from the detection of marks and traces at the scene, to their presentation in Court. R&D approached from this perspective provides no response to doubts raised by recent criminological studies, which question the effective contribution of forensic science to crime reduction, and to policing in general.Traces (i.e. forensic case data), as remnants of criminal activity are collected and used in various forms of crime monitoring and investigation. The aforementioned doubts therefore need to be addressed by expressing how information is conveyed by traces in these processes. Modelling from this standpoint expands the scope of forensic science and provides new R&D opportunities. Twelve propositions for R&D are stated in order to pave the way.     

甲醛固定及石蜡包埋组织的DNA分型及其应用

目的：建立甲醛固定及石蜡包埋组织ＤＮＡ的基因分型方法并使之应用于实际检案。方法：用Ｃｈｅｌｅｘ和有机溶剂提取法提取ＤＮＡ,以ＰＣＲ与反向探针杂交技术作ＰＭ和ＨＬＡ－ＤＱＡ１位点分型。结果：２６份包埋组织块有２４份能得到理想的分型结果。结论：本法可用于法医学个人识别和亲子鉴定     

SNPs and MALDI-TOF MS: tools for DNA typing in forensic paternity testing and anthropology

DNA markers used for individual identification in forensic sciences are based on repeat sequences in nuclear DNA and the mitochondrial DNA hypervariable regions 1 and 2. An alternative to these markers is the use of single nucleotide polymorphisms (SNPs). These have a particular advantage in the analysis of degraded or poor samples, which are often all that is available in forensics or anthropology. In order to study the potential of SNP analysis in these fields, 41 SNPs were selected on the basis of following criteria: conservation, lack of phenotypic expression, and frequency of occurrence in populations. Thirty-six autosomal SNPs were used for genotyping 21 inclusionary and 3 exclusionary paternity cases. The behavior of 5 X-chromosome SNPs was analyzed in a French representative population. Our approach to SNP typing is a multiplex PCR based amplification followed by simultaneous detection by primer extension (PEX) analyzed by Matrix Assisted Laser Desorption Ionization Time-of-Flight Mass Spectrometry (MALDI-TOF MS). The selected autosomal SNPs showed independent inheritance and gave clear results in paternity investigation. All X-SNPs were useful as both paternity and identification markers. PEX and MALDI-TOF MS, with their high sensitivity, precision and speed, gave a powerful method for forensic and anthropological exploitation of biallelic markers.     

Routine Y-STR typing in forensic casework

In the field of molecular diagnosis, forensic casework analysis is one of the most demanding investigations, due to its social impact. Optimization of DNA typing multiplex reactions with identical cycling conditions as those required by autosomal short tandem repeats (STR) multiplex reduces errors, and saves time and reagents. Previously, we validated a five Y-STRs set, all of them generating single band patterns. This work reports the optimization of combined multiplexes, a triplex (DYS19, DYS390 and DYS391) and a duplex (DYS392 and DYS393), that can be amplified in identical cycling conditions as those required by commercially available multiplex autosomal STR kits. In addition both Y chromosome multiplexes can be combined for co-injection on a capillary electrophoresis based automated sequencer. Statistical attributes of the haplotypes of the five Y-STR investigated were evaluated in unrelated males from different metropolitan areas of Argentina. This system was successfully used for investigating more than 350 forensic routine cases in our country.