PCR扩增STR位点在强奸案中的应用

利用Chelcx－100、PK、SDS、DTT对精液和阴道液、精液和血液混合班进行处理，除去女性阴道上皮细胞和血细胞获得精于细胞，提取DNA，经扩增STR位点[1]，获得高度多态性l‘ISTR位点分型图，与同一个体血液DNA的STR位点分型比较计算其概率，达到个体认定的作用．对强奸致孕案，由于STh位点等位基因按孟德尔遗传规律由亲代向子代传递，在母亲是肯定的情况下，可把STR位点分型与嫌疑父亲进行对比，计算其RCP父权相对机会）值认定嫌疑父亲为生物学父亲．通过对多起强奸案混合斑和强奸致孕案的实际应用，成功地排除或认定了罪犯，现…     

阴道液与血液中PGM_1亚型、EsD、GLOI表型相关性的研究

本文用等电聚焦电泳法和琼脂糖淀粉混合凝胶电泳法,对阴道液、血液中的PGM_1亚型、EsD、GLOI表型进行了检测。在部分妇女的阴道液中未检出其本人血液中的这些酶型,测出的比例最高的是PGM_1亚型,EsD次之,GLOI最低。凡能显示清晰酶谱带的均与同一妇女血液中的表现型相同,未见不一致的现象。同时初步探讨了月经周期和性刺激对阴道液中PGM_1酶活性的影响。     

法医物证DNA结论采纳问题

法医物证是指与人体有关的生物物证，包括案发现场存留的血液。精液、阴道分泌液、乳液、唾液、鼻涕、尿液、羊水及瘢痕、毛发、指甲、骨骼和牙齿以及人体各种组织和碎块等。通过对上述检材进行生物学检测，可起到个体识别或同一认定的作用，法医物证DNA结论的采纳，是法庭科学的难点问题，     

北京地区人群Y染色体STR遗传多态性

<正> 目前,对多数男女混合性生物检材,如血液-血液、唾液-阴道液及轮奸案中多个男性精斑混合物的法医DNA分析,仍缺乏有效的分离手段。精液和阴道液混合斑的检测虽可采用二步消化法,但亦可能分离不纯或导致男性DNA丢失[1、2]。因此,在分析混合生物样品时,只能检出各组份的混合DNA分型。由于在PCR过程中存在竞争抑制,使含量较少的DNA模板得不到有效的扩增,从而导致分型失败;即使各组份能同时检出,所得结果也常为混合基因型,使解释结果出现困难。基于上述原因,对Y染色体基因座特有的多态性研究显得更为重要[3]。     

AGCU免提取STR荧光检测试剂盒的验证

目的考察AGCU免提取STR荧光检测试剂盒．对保存在滤纸片或FTA卡上血液样本的直接扩增检测情况。方法使用人GCU免提取STR荧光检测试剂盒,对未经提取的滤纸片血液样本、FTA卡血液样本675份进行直接扩增和18个基因座的DNA分型,并对结果的可靠性进行研究。结果18个基因座检测结果与PP16和ID试剂盒分型结果一致,2000年数据库样本成功率92．3％,2001年数据库样本成功率92．6％,2004以后年数据样本及案件样本、亲子鉴定样本成功率在99％以上。结论AGCU试剂盒可以成功地对滤纸片、FTA卡样本的18个STR基因座进行直接扩增检测,检验结果稳定,分型准确。     

中国人精子黄递酶Ⅲ型的分布及精斑黄递酶Ⅲ型的检出

应用超薄层聚丙烯酰胺凝胶等电聚焦技术对辽宁地区200例正常人精子黄递酶Ⅲ型(DIA)的分布进行了研究。其基因频率分别为DIA_3~1=0.6850,DIA_3~2=0.2875,DIA_5~3=0.0275。由阴道液、唾液、乳汁及无精子症精液不能判定DIA_3型。室温保存精斑可检出时限为9个月。     

人精液γ-谷氨酰转肽酶（GGT）多态性研究

γ-谷氨酰转肽酶(GGT EC2,3,2,2,)存在于人体许多组织之中,它催化γ-谷氨酰基因的转移。七十年代中期,国外学者发现人类精液和阴道分泌液中存在高活性的 GGT,并具有多态性和稳定性。目前,国内尚未见有人类精液 GGT 多态性的分型报导,本文应用平板圆盘电泳技术,分析研究中国人群中精液 GGT 的多态性,现将初步研究结果报告如下。     

免疫磁珠法分离混合样本中血液成分的探究

目的 基于免疫磁珠法分离脱落上皮细胞中的白细胞，消除或减弱混合检材中血液来源STR分型对结果分析的干扰。方法 分别取两名不同个体的血液和口腔脱落上皮细胞，按不同比例制备成混合样本作为实验组，并同时制备细胞量相等的对照组。应用免疫磁珠法分离实验组样本中白细胞，并与对照组以相同条件进行DNA提取、扩增、分型，对比两组STR分型结果。结果 当血液量较少时，对照组为混合STR分型，经免疫磁珠法分离混合检材中白细胞后，实验组为单一来源STR分型；当血液量较多时，此时对照组为混合STR分型，但口腔脱落上皮细胞来源的STR分型谱带峰高较低甚至丢失，经免疫磁珠法后，实验组仍为STR混合分型，但口腔脱落上皮细胞来源的STR分型谱带成为主峰；当混合样本中口腔脱落上皮细胞微量，血液占比极大时，此时对照组为血液来源的单一STR分型结果，经免疫磁珠法后，实验组为STR混合分型，包含口腔脱落上皮细胞来源的所有等位基因分型。结论 该方法可用于分离脱落上皮细胞中血液成分，降低血液来源的DNA比例，能够改善此类混合检材中目的细胞的STR分型结果，为刑事案件中含有血迹浸染的混合生物检材的检验提供了一种新思路。     

用反相杂交技术对HLA-DQA1基因分型及其应用

目的 :HLA -DQA1其因的分型研究及在实际办案中的应用。方法 :PCR结合反相杂交检测技术对HLA -DQA1基因进行分型。结果 :检见7种基因 ,24种基因型 ,获得上海地区HLA -DQA1基因的基因频率及基因型的频率分布数据。结论 :对法医学中常见的血液 (痕 )、牙齿、精斑、混合斑、人体组织等检材进行HLA -DQA1分型检测 ,其结果稳定可靠 ,为实际办案提供了科学依据     

PCR－RFLP技术鉴定ABO基因型的法医学应用研究

目的建立PCR－RFLP、非变性PAG胶垂直电泳和银染技术进行ABO基因分型的方法体系,并对200名广东汉族人群ABO基因型频率进行了调查。方法用Chelex－100和酚、氯仿抽提法处理样本,PCR扩增后用非变性聚丙烯酰胺凝胶垂直电泳和银染技术检测分型。结果ABO位点特异性扩增片段长度为175bp～210bp,6种基因型频率分布为0.0250～0.4300,杂合度H值为0.5162,个体识别力DP值为0.7111。结论该方法可成功运用于血液、血痕、精斑、毛发、骨组织和混合斑等检材的个体识别及亲权鉴定的检验。     

DIA3 phenotyping in human tissues, dental pulps, and hair roots by isoelectric focusing

The polymorphism of DIA3 was investigated in tissues of various human organs, dental pulps, and hair roots by isoelectric focusing. DIA3 types were demonstrated from tissues of brain, prostate, testis, ovary, and uterus, but not from tissues of spleen, pancreas, heart, liver, muscle, lung, skin, and kidney. Determination was possible from dental pulps stored at room temperature for up to 2 weeks and from fresh hair roots. The results show that the DIA3 typing by isoelectric focusing is useful for medicolegal individualization of brain, reproductive organs, teeth, and hairs.     

DIA3 phenotyping in human semen and seminal stains by isoelectric focusing

The polymorphism of DIA3 was investigated by isoelectric focusing in semen samples from 235 unrelated Japanese volunteers and patients. Besides the three common phenotypes seven samples of the type 3-1 were observed. However, readable isoenzyme patterns were not demonstrated in semen samples of oligospermia under about 10 X 10(6)/ml sperm cells. The allele frequencies were DIA3*1 = 0.821, DIA3*2 = 0.164, and DIA3*3 = 0.015. The DIA3*1 frequency in oligospermia (0.765) was lower than that in normospermia (0.836). The isoelectric focusing method was successfully applied to phenotyping DIA3 in seminal stains; each phenotype was demonstrated at 37 degrees C for up to 4 weeks, at room temperature for up to 8 weeks, and at 4 degrees C for over 12 weeks after stain formation. In vaginal swabs the isoenzyme bands were very faint and not identifiable.     

人与动物毛发角蛋白组分的等电聚焦图谱分析

本文用聚丙烯酰胺等电聚焦(PAGIEF)对来自6个目、15个科、20种动物毛发角蛋白组分进行了研究。发现人与动物毛发角蛋白组分的PAGIEF谱型有显著差异;不同种属的动物毛发角蛋白组分的PAGIEF谱型也互不相同,它们各自都有独特的角蛋白PAGIEF谱型。作者认为,本方法可用于鉴定毛发的种属。     

Typing study of human semen DIA3 by isoelectric focusing--distribution in the Wuhan population, China

Human semen DIA3 typing was studied by isoelectric focusing on ultra-thin-layer polyacrylamide gel which resulted in a simpler and more definite separation of the products of DIA3 alleles than hitherto. In 198 semen samples collected from unrelated Chinese males four different phenotypes were observed. The DIA3 allele frequencies were calculated: DIA 3(1) = 0.7727, DIA 3(2) = 0.2172, DIA 3(3) = 0.0101. The results of the stability study of 12 laboratory-prepared semen stains stored at room temperature suggested that DIA3 in seminal strains is a relatively stable genetic marker. Our gene frequencies have been compared to those reported in other populations.     

混合斑中精斑ABO血型的检验

<正> 1988年,壹岐裕志等报告了用吸附抗α_2-SGP 血清的硝化纤维素膜(NCF)检验混合斑中的精斑 ABO 血型,但耗时。本文作者通过对此方法的改进,采用常彩琴等研制的抗人精特异蛋白血清(anti-human seminalpeculiar protein,ASPP),采用蛋清粘片热解离法检验混合斑中精斑 ABO 血型,耗时短,效果好。现介绍如下。     

Persisting fetal microchimerism does not interfere with forensic Y-chromosome typing

Forensic Y-chromosome typing applies Y-chromosomal polymorphisms to the analysis of male/female mixed stains such as vaginal swabs in rape cases. The sensitivity of this approach exceeds that of cytological techniques combined with autosomal DNA typing. Y-chromosome typing is based on the assumption that Y-chromosomal DNA found in tissue or secretions of women must originate from a male individual, usually the perpetrator. Nevertheless, it was shown recently that fetal cells can migrate into the female body during pregnancy and can persist for decades ("persisting fetal microchimerism"). The body of a woman after a pregnancy with a male embryo can thus display a small fraction of fetal cells with Y-chromosomes. Using high sensitivity PCR protocols (reamplification with nested primers and up to 60 PCR cycles) fetal cells were previously identified in a number of maternal tissues including skin, blood, muscle and solid organs. It is, however, not clear at present, whether these cells can occur in vaginal secretions, and whether they are capable of producing false positive results in forensic Y-chromosome typing. To evaluate these questions, 66 blood samples of women with at least one son and nine vaginal swabs of women without sexual intercourse in the last 2 weeks were amplified for a stretch of the SRY gene. Eight thyroid gland tissues with already established male fetal microchimerism were used as positive control samples. Blood samples of 10 young girls without history of pregnancy were used as negative controls. Using a PCR with 10 ng of extracted DNA and 30 PCR cycles ("routine sensitivity assay") none of the samples yielded positive results. However, in a PCR with 200 ng of extracted DNA and 45 PCR cycles ("high sensibility assay"), 14% of the blood samples of mothers and 33% of the vaginal swabs amplified for SRY. Our results thus show that increasing the sensitivity of the PCR method and the amount of template DNA produce positive results while protocols used for routine Y-chromosomal typing with small amounts of DNA (approximately 10 ng of DNA) and with a limited number of PCR cycles (approximately 30) can clearly eliminate this peril.     

用酶标抗体免疫组化法测定性交后阴道内容物中精子的ABO血型

应用间接酶标抗体免疫组化法测出了53例新鲜精液、5例陈旧精斑及40例阴道分泌液中的精子与阴道脱落上皮细胞的ABO血型,30例精子与不同血型分泌型阴道分泌液孵育,未发现精子吸附阴道液中血型物质的现象,同时发现人类睾丸曲精细管中部份生精细胞、精子细胞,精子;直细精管部份上皮细胞、精液、精子;睾丸网大部份上皮细胞及副睾管中的精液与精子均含ABH抗原,故认为精子上的ABH抗原主要是精子固有抗原,13例性交后阴道内容物中精子的ABO型测定结果:7例与供者血型吻合,6例不吻合。6例中5例从O型精子中测出了女方分泌型阴道分泌液中的A或AB物质,1例B型精子未测出B及H抗原,文中对这种现象进行了讨论。     

The evidential value of peptidase A as a semen typing system

Human erythrocyte peptidase A (Pep A) displays a genetic polymorphism in blacks. Its occurrence in human semen was examined for its possible use as a semen typing system. Studies by starch gel electrophoresis, in which the Pep A was located by an improved method, were carried out on semen, semen stains, and vaginal swabs taken at known times after intercourse. In addition, a large number of vaginal swabs, negative for semen, were taken from females throughout their menstrual cycles and examined for Pep A activity. The results indicated that Pep A typing could be carried out on semen and semen stains. However, it was possible to determine the Pep A type on vaginal swabs only when they had been taken within about 3 h after intercourse.     

STR位点D19S253和D8S1179的法医学意义及应用研究

为评估STR位点D19S253和D8S1179的法医学应用价值，应用PCR和PAG垂直电泳技术对两位点的种属特异性，检测灵敏度，以及同一个体不同组织分型的同一性及不同基质和不同保存时间的斑痕分型等与法医应用有关的问题进行了研究，D19S253和D8S1179位点的检测灵敏度分别为0．25ng及0．5ng，同时两位点具有较高的种属特异性，同一性及较好重复性，且能够复合扩增，表明D19S253和D8S1179是法医学检案中较实用的两个STR标记。