联合应用多个DNA位点进行尸源鉴定

目的　研究联合应用多个DNA位点在尸源鉴定方面的应用价值。方法　应用聚合酶链反应(PCR)、聚丙烯酰胺凝胶电泳分离及银染显带的方法通过对无名尸体的有关检材与可疑双亲或子女进行亲权鉴定。结果　在 80例刑事案件尸源鉴定中 ,38例无名尸体采用较新鲜肌肉 ,通过扩增VNTR、STR多个位点得以判明尸源 ;2 9例采用腐败肌肉、 6例采用骨骼和 2例采用牙齿 ,通过扩增多个STR位点判明了尸源。仅有 1例采用腐败肌肉、 2例采用骨骼未能确定尸源。结论　运用多个位点进行亲权方法可以准确地判明尸源鉴定 ,特别对高度腐败尸体、尸块不全等情况下的尸源鉴定具有很高的实用价值     

亲子鉴定DNA检验结果分析1例

1案例资料 1.1简要案情 2011年6月15日,本市警方发现1具高度腐败无名男性尸体,为确定尸源,提取该无名尸体肋软骨、张某(嫌疑为其子)及母亲血样进行亲缘鉴定. 1.2 DNA检验 上述3份检材均采用Chelex-100法提取DNA.分别使用Sinofiler试剂盒、GoldenEye20A试剂盒和Y-filer试剂盒进行复合扩增,扩增产物用ABI-3130遗传分析仪进行电泳分析.     

短串联重复vWFⅢ扩增片段长度多态性在法医学应用的可行性研究

目的 探讨ＳＴＲ位点ｖＷＦⅢ用于法医学鉴定中的实用价值。方法 应用聚合酶链式反应（ＰＣＲ），聚丙烯酰胺凝胶电泳结合染银显带的分离方法对ＳＴＲ位点ｖＷＦⅢ扩增片断长度多态性进行研究。结果 经过优化筛选掌握了该位点的最适扩增条件，其对各类检材均获得可靠的扩增产物。本方法的灵敏度达１０ｐｇ基因组ＤＮＡ、０．５μｌ全血、１μｌ新血痕、０．１μｌ精液、０．２μｌ新鲜精斑。结论 本方法快速、灵敏、准确，在法医     

毛发DNA检验辅助认定尸源1例

1 案例资料 1．1 简要案情 2006年7月，本市某一污水沟内发现1具无名女尸（已高度腐败），提取尸体肋软骨1份送检，要求进行DNA尸源鉴定。     

高度腐败尸体指印的提取

目前对无名尸体尸源认定的方法主要有尸体相貌、DNA分析、指纹等等。当命案现场中的无名尸体高度腐败时，尸体面容完全改变，相貌辨认条件丧失；同时，高度腐败尸体细胞中的DNA已经高度降解，DNA测序大多难以得到准确结果。对于这类案件，可以通过尸体指纹与生前所留指印的同一认定来进行尸源认定，如何成功的提取高度腐败尸体的指纹是能否通过指纹进行尸源认定的关键。笔者就实际工作中所遇到的案例介绍一种教科书上未提及的高度腐败尸体指纹提取方法。     

同胞关系鉴定尸源2例

利用亲子鉴定进行尸源鉴定,国内外已很常见[1～4],当父母或单亲与子女关系已确定时,鉴定无名尸体或碎尸与其有无亲缘关系,从而达到尸源认定的目的。其中,以父母关系确定,要求鉴定无名尸体或碎尸与父母间的亲缘关系多见[2,3]。但应用该原理对其他亲缘关系(隔代成员、同胞、旁系亲属等)进行尸源鉴定并不多见,本文在实际案件中成功利用同胞血缘关系鉴定尸源2例。现报道如下:1案例资料案例1刘某(男,51岁,无业)2004年5月27日失踪。4d后,于某出租房内提取血迹及人体骨骼数根(已被煮过),案件调查认为,现场血迹及人骨可能是失踪人员刘某。由于刘某无…     

太原地区汉族人群3个STR基因座的多态性分析

短串联重复序列 (ＳＴＲ)属微卫星ＤＮＡ ,在人类基因组中广泛分布。具有ＶＮＴＲ序列特征的ＳＴＲ基因座已成为一类重要的遗传标记。ＳＴＲ基因座多态片段长度小 ,ＰＣＲ扩增成功率高 ,尤其对ＳＴＲ基因座进行复合扩增使单次扩增获得多基因座的信息量 ,简单 ,快速得到分型结果 ,在法医鉴定中具有重要的意义[1] 。应用复合扩增方法调查了太原地区汉族人群 3个ＳＴＲ即Ｄ16Ｓ539、Ｄ7Ｓ82 0和Ｐ13Ｓ317基因座的基因频率分布 ,获得了 3个ＳＴＲ多态性基因座的遗传学数据。1　材料方法1 1　实验材料1 1 1　样本　 10 1例ＥＤＴＡ抗凝血来自无亲…     

mtDNA 12S rRNA基因序列测定在腐烂动物肌肉样本鉴定中的应用

目的利用mtDNA12SrRNA基因序列测定法对宁波森林公安局查获的一例腐烂动物肌肉样本进行种属鉴定,并探讨该方法在腐烂动物肌肉样本鉴定中的应用价值。方法用酚/氯仿法从腐烂动物肌肉样本中提取出基因组总DNA,再用一通用引物通过PCR技术扩增mtDNA上12SrRNA基因的部分片段并进行序列测定。测序结果在GenBank上进行BLAST搜索,再利用DNAMAN软件进行同源性分析。结果扩增产物序列与东北虎的线粒体DNA的12SrRNA基因序列的部分片段的同源性高达99.9%。结论该动物样本为东北虎,本研究所用方法在野生动物案件的样本鉴定中有极高的应用价值。     

无名尸块的个体识别-附5例报告

在涉及无名尸体特别是无名尸块的刑事案件中,尸源认定对侦破案件有重要意义.个体识别的方法很多,大多是通过尸体个体特征、衣着、遗留物,指纹等来判断分析,随着DNA指纹技术的不断成熟,个体识别的准确性也越来越高.本文收集5例实际案件中通过法医学鉴定进行个体识别的案例,着重对法医学个体识别的方法及注意事项进行讨论,供同道参考.     

STR联合扩增鉴定碎尸案1例

聚合酶链反应（PCR）技术于1985年由美国Cetus公司的RandallSaiki、HenryErlich和KaryMullis等联合创建[1]。人类基因组DNA有3×109bp，其中10％是串联重复序列，被称为卫星DNA。卫星DNA按重复单位的长短分为大卫星、中卫星、小卫星和微卫星。其中微卫星重复单位仅由2～7hp组成，故又称为短串联重复序列（STR）。Saihi等人发现人类STR可用PCR方法扩增[2]，并且有高度多态性[3]。作者成功地利用PCR扩增PLA2A、VWA、CYP19、TH01、LPL、D6S366、D19S253、FESFPS八个STR位点对一例碎尸案中尸块做同一认定及尸源鉴定，现…     

两种DNA提取法对不同色泽肋软骨STR分型成功率的比较

目的比较两种DNA提取法对不同色泽肋软骨的DNASTR分型结果的影响。方法利用Chelex-100法和酚-氯仿法,分别对30例不同色泽的腐败尸体肋软骨进行DNA提取,STR复合扩增,ABI3100型基因分析仪对扩增产物进行检测。结果用酚-氯仿法提取的30例腐败尸体肋软骨,均检测到全部STR基因座的等位基因型。用Chelex-100法提取的肋软骨中,22例(11例白色、8例淡黄色、3例黄色)检测出全部STR基因座的等位基因型;7例(3例黄色、4例黄褐色)检测出部分STR基因座的等位基因型;1例黑灰色的腐败尸体肋软骨,未检测出STR基因座的等位基因型。结论根据肋软骨的色泽,选择适宜的DNA提取方法。对于颜色较深的肋软骨,用酚-氯仿法进行DNA提取有助于提高其STR基因座的检出率。     

Typing of deoxyribonucleic acid (DNA) extracted from compact bone from human remains.

The application of deoxyribonucleic acid (DNA) typing methods for the potential identification of unknown human remains was investigated. DNA was isolated from compact bone tissue from badly decomposed bodies and from known and unknown human remains, using a decalcification and ion wash procedure. Restriction fragment length polymorphism (RFLP) analysis of variable number of tandem repeats (VNTR) loci yielded results in some cases, but more often the DNA was too degraded to produce RFLP patterns. No RFLP profiles could be obtained from putrefied soft tissues. However, DNA extracted from compact bone tissue of human remains up to eleven years old was successfully amplified using the polymerase chain reaction (PCR) for the VNTR loci D1S80, D17S5, COL2A1, and APO B, as well as the HLA-DQ alpha locus. This is especially significant, since PCR results were obtained from those samples whose DNA had been degraded substantially and had yielded no RFLP patterns. All DNA types determined from the compact bone tissue from decomposed bodies whose identification had been established first by other means (and whose parents or offspring were available for typing) demonstrated mendelian inheritance of the alleles of the loci analyzed. These results suggest that amplification and typing of DNA extracted from compact bone of human remains could be useful in establishing the identity of a person, as well as in excluding possible false identifications.     

13个STR位点在人消化系统肿瘤组织中的变异分析

目的探讨13个CODIS-STR基因座在人消化系统肿瘤组织中的变异情况。方法收集55个个体的消化系统肿瘤组织及其正常组织和血样,Chelex100法提取DNA,用Profiler试剂盒和Cofiler试剂盒进行复合扩增,310型遗传分析仪检测。结果55例肿瘤组织中均存在细胞分裂异常现象,其中有2例肿瘤组织的STR位点发生了变异,变异的类型包括基因型改变、杂合型丢失和杂合双峰不平衡,而且变异可以是多位点同时发生。结论对肿瘤组织类型的样品进行STR分析时,应多加慎重,因为排除的位点可能来自肿瘤组织中的基因突变。     

The Achilles tendon as a DNA source for STR typing of highly decayed corpses

Forensic criminal casework often involves DNA profiling of human postmortem tissues, whereas degradational processes can affect PCR-based Short Tandem Repeat (STR) analysis. Degradation of DNA is observed to vary among different tissues and with time. Therefore, the stability of DNA in Achilles tendon samples is compared to that in muscle and kidney specimens with a variety of postmortem histories. Tissue samples from 28 autopsy cases, including 15 decomposed corpses and a control group of 13 nondecayed corpses were analysed. DNA was isolated using the All-tissue DNA Kit (GEN-IAL, Troisdorf, Germany), quantified by spectrophotometric measurement, amplified by the multiplex PCR genRES MPX-2 (Serac, Bad Homburg, Germany), and analysed on the ABI PRISM 310 Genetic Analyzer (Applied Biosystems, Darmstadt, Germany). Quantitative analysis of nondecomposed tissues revealed that the recovery of DNA was highest in kidney followed by muscle, whereas Achilles tendon tissue was the poorest source of isolated DNA. Only small amounts of DNA were present in both kidney and muscle samples from decomposed corpses. However, from decayed Achilles tendon samples twice as much DNA as from nondecayed samples could be isolated on average. These results suggest DNA to be better protected in Achilles tendons. Moreover, postmortem changes in Achilles tendons may even improve DNA isolation.     

The personal identification of many samples recovered from under the sea

An automatic and rapid DNA typing system was employed for personal identification, using fragmentary tissue samples from victims in an airplane accident. Two victims were crushed into small pieces, and 33 samples suspected to belong to them were recovered from under the sea. From each sample, 10 mg was used for testing. The parents' bloods of two presumptive victims were also examined. DNA extraction from samples was performed by the NaI method, and the obtained DNA samples were analyzed with the ABI PRISM system. Among 33 samples, 31 samples were identified to be human tissues, possibly from two victims. The other two samples seemed to be parts of marine animals. ABO blood group, STR polymorphism, and mitochondrial DNA polymorphism typing were possible in every examined human sample. Two victims' fragmentary tissues were identified by determining ABO genotype, STR type and mitochondrial DNA type. The system we employed enabled an accurate typing of many fragmentary samples in a short time, thus contributing to the fast and secure identification of many victims in such cases as big air accidents.     

Use of computer software for indirect molecular genetic identification of unidentified bodies

The authors discuss the development and use of computer software for realization of indirect DNA identification, based on identification of biological relation. Estimated algorithms of this method are based on regularities of parental signs inheritance by children and consist in comparative analysis of allele states of nuclear DNA typed locuses in unidentified bodies and probable parents of these dead subjects and subsequent estimation of the coefficients of the likelihood of hypotheses of their probable blood relationship. Available software maintain the database with identification characteristics of VNTR and STR locuses, HLA DQA1 locuses, and PM system (potential set of 23 locuses). The results of identification are presented as lists of exclusions and tables with estimated likelihood coefficients for the probability expert evaluation of relationship. The suggested computer-aided method of indirect identification is a new highly effective tool for personality identification by chromosome markers under conditions of mass information processing in examinations of unidentified corpses.     

An analysis of data curated from 5 years of identifying human remains

An archive of 5 years of cases involving the identification of human remains was curated, collecting information on: The sample type submitted, the number of STR loci yielding interpretable results, the kinship challenge posed, and the outcome for the case. A total of 129 cases of remains ID were investigated using manual DNA extraction and recovery methods with amplification of STR markers using the Power Plex 21 multiplex STR kit from Promega Corp. In 52 cases, blood spots collected by the ME were provided as sample and in 100% of those cases, probabilities of relatedness to the reference samples was ≥99%. In 77 cases, tissue other than blood was provided as a source of DNA. These other samples were grouped categorically into long bones (femur and tibia; 40 cases), skull bones/teeth (11 cases), other bones (16 cases), and tissue (normally adherent to bone) (10 cases). Reference samples provided for cases included alleged parents or child(ren) of the victim (86 cases), alleged full siblings of the victim (38 cases), or alleged second-order relatives (five cases). The overall success rate in confirming the identity of the source of the remains in these cases was 89.2%. Our results demonstrate that a laboratory can be often successful identifying human remains using methods easily implemented in any DNA typing laboratory.     

汉族12个STR位点群体遗传学调查

目的240个汉族无关个体12个STR位点基因频率调查及其法医学应用;方法采用12位点复合扩增及变性聚丙烯酰胺凝胶电泳基因分型;结果该系统12个STR基因位点在汉族人群中均为高识别率位点,特别适合于陈旧血痕检验;结论12位点STR-PCR复合扩增系统检测方法简便,经济实用,在法医个体识别和亲子鉴定中具有应用价值。     

Results of collaborative study regarding the standardization of the Y-linked STR system DYS385 by the European DNA Profiling (EDNAP) group.

Y-chromosome linked short tandem repeat (STR) loci are inherited as a closely linked haplotype, which appears to remain stable in a given paternal lineage over many generations. In forensic cases, Y-linked STRs are particularly useful for the identification of human remains as well as in rape cases with mixed male/female stain samples. DYS385 is derived from tandemly duplicated segments of the Y chromosome thus giving rise to two fragments of variable length which do not behave like alleles but genotypes. The European DNA Profiling (EDNAP) group has carried out a collaborative exercise among 14 participating laboratories using DYS385 for typing of five unknown bloodstains and a control sample. Furthermore, population data from eight different European countries with samples sizes between 91 and 150 male individuals were collected. The results confirm previous observations that DYS385 is one of the most informative Y-linked STR loci. It could also be demonstrated that reproducible results can be obtained independently from the electrophoretic separation and detection methods used. Thus DYS385 may serve as a useful complementation to the routinely used autosomal STR systems in special cases.