应用ABC—ELISA进行人血痕种属鉴识的研究

本文报道了用生物素亲和素复合物—酶联免疫吸附测定(ABC—ELISA)点斑法对不同种动物血痕及人血痕进行人种属鉴识的实验结果。反应在硝酸纤维素薄膜上进行,结果可直接用肉眼判别,也可经透明处理后由酶标比色计判读。作者认为,本法可提高人种属鉴识灵敏度。本法也可适用于法医物证检验中相似的免疫测定。     

体外DNA扩增技术鉴定血痕性别三例报告

本文报道用体外DNA扩增技术对3例刑事案中的人血痕标本进行性别鉴定。其方法是从血痕标本中微量抽提DNA,用蛋白酶K进行消化。然后用两组引物Y1.1与Y1.2和Alu9.1与Alu9.2及国产FD耐热DNA聚合酶进行聚合酶链反(PCR)扩增DNA,电泳分析Y及Alu重复序列,从而判断血痕的性别。     

PCR扩增ZFY/ZFX基因鉴定人类干血痕性别

应用 PCR 技术同时扩增人 ZFY 和 ZFX 基因特异的 DNA 序列,在男性血痕中可检测到两种扩增产物,即340bp 长的 ZFY 基因及488bp 长的 ZFX 基因特异 DNA 片段;在女性血痕中仅可检测到488bp 长的 ZFX 基因特异 DNA 片段,据此判定干血痕性别。干血痕的最小检出需要量为0.125μl 血液量的血痕。室温保存10年的血痕可以准确判定性别。ZFY 基因位于 Y 染色体短臂。本方法同时检测两条性染色体,可以避免由于扩增失败或 Y 染色体长臂变异出现的假阴性或假阳性。扩增产物经琼脂糖凝胶电泳即可区分。     

用引物Y_3,Y_4和DNA扩增技术作血液(痕)和毛根性别鉴定的研究

作者用引物Y_3、Y_4和DNA聚合酶链式反应(PCR)作微量人类血液(痕)和毛根的性别鉴定。扩增的靶序列位于Y染色体DNA特异3.4kb重复序列中,扩增产物为460bp。检材用量为:新鲜血液0.5μl、血痕纱纤维1mm、毛根单个。20例保存4个月的血痕与2例保存6年半的血痕性别判定结果均正确,无性别记载的保存9～11年的3例血痕显现了清晰的460bpY特异DNA扩增带。15例保存20天的自然脱落毛根性别判定结果均正确。本法省略了检材处理中的酚-氯仿抽提DNA等纯化步骤,既简化了实验操作,又减少了检验过程中外源DNA的污染机会和样品DNA的损耗,使这一性别鉴定方法更符合法医学实践的需要。     

法医DNA分型的可重复性

为了了解国内法医DNA分型的可靠性，选择酪氨酸羟化酶基因第1内含子什＂mantyrosinehydroxylasegene,intron1）的短串联重复序列（STR）TH01基因座为遗传标记，在国内5个法医实验室进行了DNA分型的可重复性研究，报道如下。材料与方法一、样本制备和分送国内5个法医实验室参加本次研究工作，由华西医科大学法医学系作为协调单位，负责制备与分送盲测样本。每个法医实验室4份样本，2份为纯化DNA，2份为血痕。血液样品采自成都地区无血缘关系汉族个体，常规方法提取DNA；血痕样本由另外两名个体新鲜血液0．5ml滴于滤纸上制成，室温晾干…     

血痕识凶

"110吗?这里有打斗的痕迹,地上和墙上有血痕。我怀疑这里曾经发生过凶杀案。你们是不是能派人过来查看一下。我现在的位置是……"110报警中心有时会接到这样的报警电话。电话中提到的血痕是法医实践中最常见的生物物证,是法医物证检验的主要对象之一。警方抵达疑似案发现场后,会对血痕进行搜集和检验,首先要确定是否是真的血痕,接着要判断血痕是人留下的还是动物留下的。在确定是人的血痕之后,通过科学的检验方法,可能判断出留下血痕者的真实身份,为侦破刑事案件提供十分有用的线索。     

用引物Y_3、Y_4和PCR方法鉴定性别的法医学应用

用Y3、Y4和Alu9．1、Alug．2两对引物和PCR方法检测陈旧血痕和毛根的性别获得成功。引物Y3、Y4扩增的靶序列位于Y染色体特异3．4Kb重复序列中，扩增产物为460bp；引物Alu9．1、Alu9.2用以扩增男女共有的Alu重复序列，扩增产物为130bP。室温保存13年之久的19例脐带血血痕（男性9冽，女性10例）和室温保存10～11个月的10例已知性别自然脱落毛根（男性6例，女性4例）的性别测定结果均正确；对一起凶杀案的血痕性别测定为定案提供了重要证据。本方法简化了样品的前处理过程。     

尼龙植绒拭子与棉拭子血痕DNA分型效果比较

目的探讨尼龙植绒拭子与棉拭子血痕DNA分型检验的效果。方法取抗凝全血用去离子水倍量稀释2、4、8、16、32、64、128倍,各取1μL分别滴加于尼龙植绒拭子与棉拭子头部,干燥制成血痕,应用Chelex-100提取每个浓度两种材质拭子的血痕样本DNA,采用Identifiler Plus试剂盒进行复合扩增,3500XL型遗传分析仪进行检测,对比各组DNA分型检验成功率。结果 1检验成功率:稀释16~64倍尼龙植绒拭子血痕均明显高于相同稀释倍数的棉拭子血痕(P0.001);其余浓度血痕两种材质拭子的检验成功率无明显差异;2分型图谱峰高和均衡性:不同浓度尼龙植绒拭子血痕峰高多为棉拭子血痕的2倍以上,16~64倍稀释血痕尼龙植绒拭子均衡性更好。结论尼龙植绒血痕拭子的检验效果优于棉拭子血痕,在微量血痕检验时优势更明显。     

Y染色体法医DNA检验策略

法医DNA检验在实际工作中发挥了重要作用,其中针对Y染色体进行的DNA检验,可以开展家系排查和辅助父系亲缘鉴定,为案件侦查提供重要线索。本文针对Y染色体DNA检验,讨论完整利用染色体具有的信息,制定整体检验策略,以期为相关研究和试剂盒开发研制提供参考。     

对420例血痕性别测定的研究

<正> 本文收集420例干血痕,男性364例,女性56例,经制片、国产盐酸阿的平染色,用奥林巴斯研究显微镜,对血痕Y荧光小体进行了观测,结果364例男性血痕的Y小体检出率平均为29.69％,其变动范围在9～63％;56例女性血痕的类Y小体平均检出率为0.75％,其变动范围0～4％;实验的同时,对150例男性血痕和25例女性血痕涂片,运用透     

Rapid quantification and sex determination of forensic evidence materials

DNA quantification of forensic evidence is very valuable for an optimal use of the available biological material. Moreover, sex determination is of great importance as additional information in criminal investigations as well as in identification of missing persons, no suspect cases, and ancient DNA studies. While routine forensic DNA analysis based on short tandem repeat markers includes a marker for sex determination, analysis of samples containing scarce amounts of DNA is often based on mitochondrial DNA, and sex determination is not performed. In order to allow quantification and simultaneous sex determination on minute amounts of DNA, an assay based on real-time PCR analysis of a marker within the human amelogenin gene has been developed. The sex determination is based on melting curve analysis, while an externally standardized kinetic analysis allows quantification of the nuclear DNA copy number in the sample. This real-time DNA quantification assay has proven to be highly sensitive, enabling quantification of single DNA copies. Although certain limitations were apparent, the system is a rapid, cost-effective, and flexible assay for analysis of forensic casework samples.     

A study of sex identification of trace, dried bloodstains using a Y-chromosome-specific deoxyribonucleic acid (DNA) probe

A new method is discussed which examines trace, dried bloodstains by gel in situ hybridization using a Y-chromosome-specific deoxyribonucleic acid (DNA) probe to determine the sex of the bloodstain for forensic medicine application. The complete DNA is transferred directly by electrophoresis onto the gel intact, bypassing the possibilities of impurities contaminating the sample and of DNA degradation. The method has proven accurate for small (2.5-mm-diameter) samples aged up to eight years and is quick, simple, and easily read.     

Sex determination of blood stains with a recombinant DNA probe: comparison with radioactive and non-radioactive labeling methods

A recombinant DNA probe hybridizing specifically to human repeat DNA sequence (pHY10) of which about 3000 copies are present on the Y chromosome was used for sex determination of degraded DNA samples of blood stains. Human blood stains of male and female origin were readily differentiated with the pHY10 DNA probe. This radioactive technique enabled reliable and sensitive sex determination from blood or dried blood stains greater than 20 years old. Less than 1 microliter of blood or 1 piece of 0.5 cm length thread of blood stain from cotton fabric was sufficient for the test using dot blot hybridization. Compared with the radioactive labeling method, the photobiotin labeling method showed one thirtieth to one fiftieth lower sensitivity and presented some problems which are expected to be resolvable.     

A simple and cost-effective method for preparing DNA from the hard tooth tissue, and its use in polymerase chain reaction amplification of amelogenin gene segment for sex determination in an Indian population

The use of teeth as an important resource in the analysis of forensic case history by polymerase chain reaction (PCR) or other related methods has been reported. However, a major drawback in using teeth has been that the DNA is present only in trace amounts, and the methods to recover DNA from the flinty material have not been efficient or cost effective. In this report, we describe a method to prepare DNA from the hard tooth tissues. Our studies show that ultrasonication of teeth samples yields sufficient amounts of good quality DNA useful for PCR-based diagnostic methods. The teeth could serve as a reliable source of DNA for amplification-based forensic methods in sex determination. DNA could be obtained from any tooth, regardless of the age of subject. Furthermore, by using the AMEL gene-based primers in PCR, we have shown that the AMEL gene serves as a good marker for sex determination in the Indian population. In our study, the PCR-based method was sensitive and proved to be successful for sex determination with a complete specificity.     

Sex identification of bloodstains by radioimmunoassay of sex hormones

A forensic application is reported for the sex determination of subjects whose dried bloodstains are analyzed by radioimmunoassay for testosterone and progesterone. Blood specimens of ten males and 15 females were collected, prepared as bloodstains, and then assayed at four-different time intervals for testosterone (T) and progesterone (P) contents up to 3 months later. The ratio of the two hormone contents ($\text{P}\text{T}$) was used to establish the sex origin of the dried blood specimens.     

Human identification and sex determination of dental pulp, bone marrow and blood stains with a recombinant DNA probe

Recombinant DNA hybridizing specifically to a 300 nucleotide repeat DNA sequence (BLUR8) of human specificity and to human repeat DNA sequence (pHY10) on the Y chromosome was used for human identification and sex determination of degraded DNA samples of blood stains, dental pulp, and bone marrow. This radioactive technique enabled reliable and sensitive human and sex determination from blood stains that were more than 80 years old. Less than 1 piece of 0.5 cm length thread of blood stain was enough for both tests. DNA from relatively fresh dental pulp and bone marrow was clearly identified. The human identification test, which could recognize up to 0.3 ng DNA correctly, was 3 to 5 times more sensitive than the sex determination test.     

用PCR扩增X-Y-特异片段性别鉴定的研究

采用5%Chelex-100处理方法提取 DNA,用 PCR 扩增 Amelogenin 基因片段检测血痕和毛根的性别。扩增一个样品只需1/10体积3mm~2血痕 Chelex-100处理液或1/10体积一根毛发 Chelex-100处理液(含模板 DNA)。100例血液样品结果显示,在男性可同时显现977bpX 特异片段和788bpY 特异片段,而在女性只观察到977bp X 特异片段,该方法用于性别鉴定可靠、灵敏、方便。     

Metric Sex Determination of the Human Coxal Bone on a Virtual Sample using Decision Trees

Decision trees provide an alternative to multivariate discriminant analysis, which is still the most commonly used in anthropometric studies. Our study analyzed the metric characterization of a recent virtual sample of 113 coxal bones using decision trees for sex determination. From 17 osteometric type I landmarks, a dataset was built with five classic distances traditionally reported in the literature and six new distances selected using the two-step ratio method. A ten-fold cross-validation was performed, and a decision tree was established on two subsamples (training and test sets). The decision tree established on the training set included three nodes and its application to the test set correctly classified 92% of individuals. This percentage was similar to the data of the literature. The usefulness of decision trees has been demonstrated in numerous fields. They have been already used in sex determination, body mass prediction, and ancestry estimation. This study shows another use of decision trees enabling simple and accurate sex determination.     

Discriminating sex in South African blacks using patella dimensions

For many years, sex determination has been carried out on skeletal remains to identify individuals in forensic cases and to assess populations in archaeological cases. Since it has been shown that not all bones are found in a forensic case, discriminant function equations should be derived for all bones of the body to assist in sex determination. Numerous studies have shown the usefulness of bones of the lower extremity (e.g. femur, tibia) in sex determination using discriminant function analysis, but the use of patella measurements has not been extensively investigated for this purpose. It is therefore the aim of this study to derive discriminant function equations for sex determination from measurements of the patella of South African blacks as represented in the Raymond A. Dart Collection of Human Skeletons. A total sample of 120 (60 male, 60 female) patellae were measured using six measurements. The Statistical Product and Service Solutions (SPSS) program was used to derive the equations. Stepwise and direct analyses were performed with the highest rate of classification of 85% thereby making the patella useful for sex determination. Thus, the proposed equations derived from this study should be used with caution and only on the South African black population group.