应用AFLP检测大麻遗传多样性

目的 筛选对于大麻多态性好的AFLP引物标记来区分大麻品种.方法 利用AFLP技术用55对引物组合对12个大麻地域品种进行初筛.结果 选出5对多态性好的引物组合进行了遗传多样性研究.每对AFLP引物组合扩增出47～76务带.共获得285条带,其中多态性条带为99条以及10条品种特异带.结论 AFLP对于大麻具有很高的分辨率,为今后深入地研究大麻植物遗传多样性奠定了很好的基础.     

植物物证的法医学应用

随着植物物证在案件侦破过程中的作用日益明显,应用植物学研究方法为刑事案件提供线索、证据的“法医植物学”越来越受到关注。法医植物学常用的实验技术有形态学检验、理化检验、分子遗传学检验等。本文通过整理法医植物学研究的相关文献及案例,以列举的形式简要阐述不同检验方法的利弊,总结法医植物学应用中需要注意的问题,供案件现场勘验时参考。     

DNA分子标记技术在法医植物学中的应用

法医植物学是一门研究与法律事件相关的植物证据的科学。植物物证的DNA分子标记技术是近年来法医植物学研究的主要方向。本文系统地综述了目前应用于法医植物学中的各种DNA分子标记技术，对这些方法的实际应用案例以及可能应用方向进行列举，最后总结出这些分子生物学技术相较于传统植物形态分类方法的优点。     

等位基因特异性PCR技术及其法医学应用

等位基因特异性PCR（allele-specific polymerase chain reaction,AS-PCR）是一种基于等位基因特异性引物引导的PCR技术,可以有效地分析单核苷酸多态性（SNP）,包括碱基的转换、颠换以及插入/缺失多态性,在疾病研究、分子诊断以及法医物证学研究中具有很好的应用价值.本文系统地综述了AS-PCR技术的原理、检测手段、改进方法及在常染色体、Y染色体和线粒体SNP等领域的研究成果,探讨其法医学应用价值.     

第19届国际法医遗传学会议论文简介

<正> 2001年8月29日～9月2日,在德国明斯特(Moster)召开了第19届国际法医遗传学会。此会议共收到来自不同国家的263篇论文,主要归纳为:(1)SNP 的复合扩增及其高通量检测技术的研究;(2)常染色体 STR 基因座复合扩增多态性及群体遗传学数据;(3)线粒体 DNA 序列多态性及其单倍型群体频率分布数据;(4)性染色体(X 和 Y)STR 基因座的复合扩增多态性和突变的研究。SNP 具有密度高、遗传稳定、分析易自动化等特点,在整个生物学界掀起了基因多态性研究的新热潮,同时为法医     

大麻的DNA分析检验技术

本文简要回顾了大麻的一些常规检验方法,并重点综述了大麻植物的DNA分析检验技术,包括随机扩增多态性DNA(RAPD)、测序扩增区段(SCAR)、短串联重复序列(STR)、扩增片段长度多态性(AFLP)、单核苷酸多态性(SNP)等DNA分子标记检测法。并且对这几种技术的应用范围及前景做了简要分析,以期为实践工作提供基础理论帮助。     

法医DNA分型

编者按DNA(脱氧核糖核酸)是生物遗传物质.DNA分型技术在法医物证鉴定中应用日益广泛,在各类重大 刑事案件的侦破中发挥了越来越重要的作用.近10多年,是法医学发展最快的时期,DNA分型技术跨了两大步: 1985年建立的第一代分型技术--DNA指纹,实现了物证鉴定从否定到认定的历史性转变;进入90年代,PCR(聚 合酶链反应)技术问世,可以在体外完成扩增复制靶DNA片段的全过程,是生命科学一项划世纪的进展.以PCR为 基础的第二代DNA分型技术在检测灵敏度上有重大突破,解决了半个多世纪以来困扰法医的微量、陈旧、腐败检材 鉴定的难题.DNA分型的高科技,以其无可争议的认定或否定的鉴定结论,为法庭提供确凿的证据,成为打击犯罪 的有力武器.从本期起,对<法医DNA分型>进行专题讲座,将分期介绍DNA多态性基本原理,DNA分型结论的 评估,法医DNA分型技术与方法,以及DNA分析技术展望等四个部分,以飨读者. 许多涉及人体伤害、死亡的刑事案件,法医常要对现场 遗留的血痕、精斑、毛发、骨骼等人体材料作出鉴定,分析 这类物证是来自被害人或罪犯,称为个人识别或同一性鉴定. 个人识别或同一性鉴定可以通过检测血型进行判断:血型不 同,可排除同一性;血型相同,则不排除同一性.血型如同 人的一种特殊标记,每一个人有独自的血型.血型严格按照 一定的遗传规律从亲代遗传给子代,故称血型是人类的一种 遗传标记.通过对遗传标记的检测分析,人类可分成若干类 型.这种现象叫作遗传学多态性.如按ABO血型分类,就有 A、B、AB和O等4类.多态性的产生原因,是基因座上的 碱基发生了点突变,形成等位基因.在DNA分子中,含有大 量的高多态性基因座及遗传标记,应用分子生物学方法直接 检测分析DNA遗传标记的型别,实现个人识别的目的.通过 分析亲代和子代的遗传标记作亲子鉴定,可以判断父母和孩 子是否亲生关系.对遗传学多态性和遗传标记的研究,为法 医学个人识别和亲子鉴定提供了理论基础.     

基于下一代测序的全解析度STR分型研究进展与展望

下一代测序技术具有高通量、高速度、集成化、低成本等显著优势,近年来已在科研和临床诊断领域得到广泛应用,在法医遗传学领域亦具有重要应用前景。当前主流的STR分型方法仅关注序列的长度多态性,然而由于核心重复结构存在差异或扩增区段内存在SNP,序列长度相等的等位基因可能是具有遗传稳定性的完全不同的等位基因,此类STR序列多态性是个体识别或亲缘关系分析的宝贵资源。基于下一代测序的STR分型在现有数据输出方式基础上,允许进一步关注STR的序列多态性,对STR基因座进行全解析度分型,显著提升STR基因座的个体识别能力。本文以法医STR遗传标记和下一代测序技术为关注焦点,系统综述基于下一代测序的全解析度STR分型领域国际最新研究进展,深入探讨该技术在法医DNA实验室的实际应用潜力和可能面临的挑战,希冀对相关研究和实践提供参考。     

X染色体遗传标记研究进展

随着人类基因组计划的进展,全面深入地了解个体和群体间基因组的变异或多态性已成为可能,并日益显示其重要性。X染色体遗传标记的遗传多态性较多地表现在短串联重复序列和单核苷酸的多态性。这类多态性有助于解释个体的表型差异、不同群体和个体对疾病,特别是对复杂疾病的易感性、以及对各种药物的耐受性和对环境因子的反应,因此受到临床及法医工作者的重视。本文收集了国内外X染色体遗传标记在临床疾病诊断及法医DNA工作者实践中的研究和应用资料,汇总报道了X染色体遗传标记的研究进展。     

人血清、血痕中HLA——A_2和HLA——B_(40)抗原的测定

具有复杂的遗传多态性的HLA(人类白细胞抗原)系统的实际应用正日益受到重视.在医学临床上,建立在HLA分型基础上的有关输血、器官移植、卵性诊断、疾病关联等研究和应用规模正日趋扩大和深入;在法医实践中,利用HLA分型进行亲子鉴定的工作也业已在国内开展. HLA的多态性,显现了其在作个人识别时具有极高的排除率.由于HLA不仅以     

Analysis of human specificity in AFLP systems APOB,PAH, and D1S80

We have previously characterized and databased three human amplified fragment length polymorphism (AFLP) loci: the hypervariable regions 3′ to apolipoprotein B (APOB), phenylalanine hydroxylase (PAH) and at locus D1S80. The analysis utilized polymerase chain reaction (PCR) technology for human identification in forensic and paternity testing. This study extended that work by assessment of specificity of amplicons produced with non-human and human control DMAs for APOB, PAH and D1S80 under high and low stringency PCR conditions. It was seen that primate and other animal templates (with the exception of chimpanzee) yielded products below the human allele range under high stringency PCR parameters. Under reduced stringency PCR with animal and primate samples, reproducible genetic fingerprints were generated spanning the human allele range. The patterns were produced with defined human AFLP primer pairs under specifically relaxed PCR reaction and thermalcycling parameters. They showed genetic relationships between species at the DNA level. Amplicon patterns were compared for band size and intensity matches within the PCR synthesis range defined by the conditions used. This technique could become a useful tool in species identification and molecular evolutionary studies.     

Genetic variation in hemp and marijuana (Cannabis sativa L.) according to amplified fragment length polymorphisms

Cannabis sativa L. (Cannabaceae) is one of the earliest known cultivated plants and is important in the global economy today as a licit and an illicit crop. Molecular markers distinguishing licit and illicit cultivars have forensic utility, but no direct comparison of hemp and marijuana amplified fragment length polymorphism (AFLP) has been made to date. Genetic variation was surveyed in three populations of fiber hemp and a potent cultivar of marijuana using AFLP markers. Ten primer pairs yielded 1206 bands, of which 88% were polymorphic. Eighteen bands represented fixed differences between all fiber populations and the drug cultivar. These markers have practical utility for (1) establishing conspiracy in the cultivation and distribution of marijuana, (2) identifying geographic sources of seized drugs, and (3) discriminating illegal, potent marijuana cultivars from hemp where the cultivation of industrial hemp is permitted.     

Identification of Acer rubrum using amplified fragment length polymorphism

Amplified fragment length polymorphism (AFLP) analysis of botanical forensic evidence provides a means of obtaining a reproducible DNA profile in a relatively short period of time in species for which no sequence information is available. AFLP profiles were obtained for 40 Acer rubrum trees. Leaf material from five additional species was also typed. Genomic DNA was isolated using the DNeasy Plant Miniprep Kit (Qiagen, Valencia, CA), double-digested by two restriction endonucleases (EcoRI and MseI) and ligated to oligonucleotide adapters. Two consecutive PCR reactions (pre-amplification and selective amplification) were performed using a modification of the AFLP protocol described by Gibco (Invitrogen, Rockville, MD). The DNA fragments were separated by capillary electrophoresis using the CEQ 8000 DNA Fragment Analyzer. A number of Acer rubrum species-specific peaks were identified. In addition, within this closed set of samples, 15 of 16 (93.8%) blind samples were correctly identified. AFLP data can be used to determine the species of botanical evidence or to associate a sample to a source. This information can be used in forensic investigations to link a piece of evidence with a particular location or suspect.     

二代测序技术在Y染色体遗传标记分型中的法医学应用

法医遗传学领域常利用Y染色体的父系遗传特点,对非重组区遗传标记进行检测并用于亲缘关系鉴定、混合斑检测、家系排查以及种族推断等研究。目前毛细管电泳仍是应用最为广泛的检测技术,基于该技术的商业化检测试剂盒及数据分析处理系统十分成熟。随着生物信息量的增长,传统检测技术通量低的弊端逐渐显现,推动了法医DNA分型技术的革新。近年来,二代测序(next generation sequencing,NGS)技术发展迅速,其应用已被推广到包括法医遗传学在内的各领域,为Y染色体遗传标记的检测提供了新的技术手段。本文就NGS技术应用于法医学Y染色体遗传标记检测的研究现况和应用前景进行阐述,以期为后续司法实务提供新思路。     

A new approach to detect a set of SNP-SNP markers: Combining ARMS-PCR with SNaPshot technology

Microhaplotypes have become a new promising forensic genetic marker in recent years. The microhaplotype composed of two SNPs, SNP-SNP, indicates strong application potential because of the shortest fragment and good polymorphism and without the interference of stutter and high mutation rate as short tandem repeats (STR) and low polymorphism as a single SNP. Currently, the most common method to detect microhaplotypes is massively parallel sequencing (MPS), however its high cost and the need for special instruments limit its use in general forensic laboratories. In this study, we screened out 8 new SNP-SNP loci and established a new detection method by associating multiplex ARMS-PCR and SNaPshot technology. Firstly, we introduced ARMS-based PCR for SNP1. Then, SBE primers for SNaPshot assay were designed as 20–25 bp upstream complementary sequence next to the position of SNP2. Finally, 8 loci were built into one panel based on different SBE primer lengths and fluorescence colors. In brief, by combing ARMS-PCR and SNaPshot technology, it is easy and fast to profile the SNP1 and SNP2 orderly of the SNP-SNP microhaplotype based on CE platform. Our results suggested that the 8 loci have relatively high polymorphism as well as robust performance.     

利用荧光AFLP技术检测罂粟DNA多态性

目的 利用荧光AFLP检测技术检测罂粟植物DNA多态性。方法 用Axygen公司的AxyPrep DNA试剂盒提取了12株产于缅甸和中国云南省昆明市宜良县罂粟植株的DNA，用Eco RI和Mse I对总DNA进行酶切。连接人工接头，预扩增和选择性扩增，其产物在CEQ8000遗传分析系统上检测。结果 64对选择性扩增引物中8对引物能得到20条以上的扩增片断，具有高度多态性。结论 荧光AFLP技术可用于罂粟DNA多态性的检测。     

DNA polymorphism detection of Papaver somniferum L using fluorescent amplified fragment length polymorphism

OBJECTIVE: To detect DNA polymorphism of Papaver somniferum L using fluorescent Amplified Fragment Length Polymorphism. METHODS: Genomic DNA was isolated using the AxyPrep DNA Kit, double-digested by two restrictional endonucleases (Eco RI and Mse I) and ligated to oligonucleotide adapters. After Pre-amplification and selective amplification, the DNA fragments were separated by capillary electrophoresis using the CEQ8000 DNA Fragment Analyzer. RESULTS: More than 20 fragments of highly polymorphic products were obtained by 8 pairs of primer from 64 selective amplifying primer pairs. CONCLUSION: The fluorescent AFLP technique can be used to detect the DNA polymorphism of Papaver somniferum.     

A simple DNA extraction method for marijuana samples used in amplified fragment length polymorphism (AFLP) analysis

As a first step in developing a molecular method for the individualization of marijuana samples, we evaluated a plant DNA extraction kit. The QIAGEN plant DNeasy method uses a spin column format for recovery of DNA and is effective for obtaining high molecular weight DNA from leaf, flower (bud), and seed samples of marijuana. The average DNA yield was 125-500 ng per 100 milligrams of fresh plant tissue. The recovered DNA was of polymerase chain reaction (PCR) quality as measured by the ability to generate reproducible amplified fragment length polymorphism (AFLP) profiles. AFLP is a technique used to create a DNA profile for plant varieties and is being applied to marijuana samples by the authors to link growers and distributors of clonal material. The QIAGEN plant DNeasy method was simple, efficient, and reproducible for processing small quantities of marijuana into DNA.     

肋软骨分析检验技术在法医学中的应用

传统法医人类学对肋软骨的分析检验仅限于形态学检验.近年来,法医影像学以及分子遗传学的发展丰富了人类肋软骨的分析检验技术,并且已经能够运用于法医学实践中.本文从肋软骨的组织学特性入手,综述了近年来肋软骨在法医人类学性别判定、年龄推断等方面的研究进展,以期为同行提供借鉴和参考.