中国黎族人群13个STR基因座的多态性调查

应用四色荧光标记引物复合扩增技术 ,对 2 13名黎族无关个体的 13个ＳＴＲ基因座进行分型研究 ,计算中国黎族 13个ＳＴＲ基因座的基因频率 ,统计分析各基因座的杂合度 (Ｈ)、多态信息总量 (ＰＩＣ)、个体识别机率 (Ｄｐ)及累积个体识别率、累积非父排除率。1　材料和方法1 1　材料黎族无关个体血样 2 13份 ,取自海南省琼中黎族自治县、陵水黎族自治县等黎族居住地。抽取静脉血液涂布于滤纸上 ,阴干保存。1 2　方法1 2 1　ＤＮＡ提取　所有检材均应用Ｃｈｅｌｅｘ 10 0法提取ＤＮＡ。1 2 2　ＰＣＲ试剂　用ＡＢＩ公司Ｐｒｏｆｉｌｅｒ Ｐｌ…     

武汉汉族人群D14S608、D16S3253、D21S1437基因座多态性

利用ＰＣＲ和ＰＡＧＥ技术 ,对武地区汉族人群Ｄ14Ｓ60 8、Ｄ16Ｓ3 2 5 3、Ｄ2 1Ｓ14 3 73个ＳＴＲ基因座进行调查。2 80例武汉地区汉族无关个体抗凝血样取自同济医院血库。ＤＮＡ提取、ＰＣＲ扩增产物检测 ,基因型命名及群体数据参照杨庆恩方法[1] ,3个ＳＴＲ基本特征和其引物参数文献[2 ] 。武汉汉族人群Ｄ 14Ｓ60 8、Ｄ 16Ｓ3 2 5 3和Ｄ2 1Ｓ14 3 7等位基因频率分布见表 1。Ｄ14Ｓ60 8、Ｄ16Ｓ3 2 5 3和Ｄ2 1Ｓ14 3 7基因座Ｈ、ＤＰ、Ｐｍ和ＰＩＣ值见表 2。　　为了从众多的ＳＴＲ位点中筛选出频率更好、分型稳定的ＳＴＲ基因座。选取了…     

直接同步扩增5种基因位点

来用直接同步ＰＣＲ扩增５种基因位点及反向打点杂交技术，使扩增产物ＤＮＡ与膜探针条杂交。再通过酶联底物显色反应对ＬＤＬＲ、ＧＹＰＡ、Ｄ７Ｓ８、Ｇｃ和ＨＢＧＧ进行联合分型。该方法特别适用于微量、陈旧或降解的ＤＮＡ样本，其操作简便、迅速，应用在亲子鉴定案例，亲子关系概率高达９９．７５％。该技术在法医学、遗传学及临床诊断中具有重要价值，并将改变ＤＮＡ分型技术难于标准化的局面．     

Chelex快速处理法成功检测肛门拭子1例

在实际办案中 ,法医检验精斑多集中于阴棉、内裤、擦拭物 ,而肛门拭子检验较少 ,笔者遇到一例 ,并成功检出犯罪嫌疑人所留精斑的基因型。2 0 0 2年 3月 ,谢某 (女 ,4 2岁 )在中山市东区某出租屋被杀。提取死者谢某阴棉、肛门拭子及嫌疑人曾某、于某、周某等人血纱 ,进行ＰＣＲ检验比对。血痕、混合斑ＤＮＡ提取见文献[1] 。ＤＮＡ扩增 :采用ＰＥ公司Ｐｒｏｆｉｌｅｒｐｌｕｓ试剂盒扩增。电泳采用 310遗传分析议电泳分型。第一次检验事主阴棉中精斑 9个ＳＴＲ位点基因型均与嫌疑人曾某相同 ,偶合概率为 1.3× 10 -12 ,从而认定精斑为曾某所…     

染色混合精斑涂片DNA检验方法的研究

建立了染色混合精斑涂片DNA的提取方法,通过两步扩增的技术与垂直聚丙烯酰胺凝胶电泳及银染色法,成功检测了3个STR位点.结果显示,所有染色涂片DNA分型均与对照血样一致,苏木素-伊红(HE)染色比酸性复红-美蓝(Baecchi)染色对DNA的降解作用要小;涂片灵敏度检验可达到少于10个精子,且室温保存一年以内的涂片其灵敏度光显著差异.该方法简单、快速、可靠,为染色混合精斑涂片的PCR分析提供了新方法.     

降解的DNA的PCR分型—电洗脱回收大片断DNA法

本文分析了降解ＤＮＡ作ＰＣＲ分型易失败的原因，提出了因小片断ＤＮＡ过多，阻碍引物与膜板的复性及阻断引物延伸的新观点。将降解的ＤＮＡ电泳，切除小片断ＤＮＡ，电洗脱回收较大片断ＤＮＡ，成功地对降解ＤＮＡ进行了ＰＣＲ分型。本方法简便、快速、有效。     

降解DNA的PCR分型-电洗脱回收大片断DNA法

本文分析了降解ＤＮＡ作ＰＣＲ分型易失败的原因，提出了因小片断ＤＮＡ过多，阻碍引物与膜机的复性及阻断引物延伸的新现点。将降解的ＤＮＡ电泳，切除小片断ＤＮＡ，电洗脱回收较大片断ＤＮＡ，成功地对降解ＤＮＡ进行了ＰＣＲ分型．本方法简便、快速、有效。     

哈尔滨(地区)汉族人群F13A01、FESFPS、vWA基因座多态性

参照美国Ｐｒｏｍｅｇａ公司Ｇｅｎｅｐｒｉｎｔ试剂盒提供的方法 ,采用Ｆ13Ａ0 1、ＦＥＳＦＰＳ、ｖＷＡ三基因座复合扩增技术 ,对哈尔滨 (地区 )汉族人群中 16 3个无关个体的 3个基因座进行基因频率分布调查 ,现报告如下。1　结果与讨论通过对 16 3名无关个体的血样经酚、氯仿萃取法提取ＤＮＡ ,ＰＣＲ扩增 ,用标准等位基因作参照物 ,电泳分离 ,银染检测后 ,进行ＤＮＡ分型和统计学分析。Ｆ13Ａ0 1基因座观察到 4个等位基因 ,9个基因型 ;ＦＥＳＦＰＳ基因座观察到6个等位基因 ,13个基因型 ;ｖＷＡ基因座观察到 8个等位基因 ,2 3个基因型 ;…     

避免生物检材PCR扩增失败的对策

ＰＣＲ是一种灵敏的ＤＮＡ检测技术，但是，在对生物学检材进行个人识别时，仍然有一定数量的检材直接ＰＣＲ扩增失败。为了有效地进行个人识别，本文分析了扩增失败的原因以及成功扩增的对策     

国外论文文摘

用聚合酶链反应法对精子阳性涂片进行HLADQ_α分型[刊,德]/Potsch L.…//德国法医学杂志,-1993,3(2),-54～58 此研究说明,以PCR法对精子阳性涂片进行HLADQ_α分型测定个体特征的可能性。以分级溶解来分离男性和女性DNA是直接在显微镜载玻片上进行的。按照DNA的萃取和扩增测定男性HLA DQ_α-型的阳性结果,至少需要20个精子。对一般使用的定影方法和染色程序以及涂片制备的实用效果进行了实验研究。以AAS载玻片用于阴道混合物的涂片比较好,因为蛋白酶K-消化后还会粘附在上面,对男性和女性细胞中的DNA能够完全分离。     

一种改良的精斑涂片DNA检验方法

目的建立一种有效的精斑涂片DNA检验方法。方法以本实验室平时积累的91例精斑涂片为研究对象,采用chelex法提取,结合硅珠法纯化浓缩,提高DNA浓度后进行PCR扩增和STR分型。结果尽管涂片上精子量较少,但chelex法的DNA提取率较高,且硅珠法可以纯化并浓缩模板DNA,本文建立的方法兼具了这两种提取方法的优点,成功率较高。结论精斑涂片可以作DNA检验,在法医检案中具有较高的应用价值。     

Cold Case Homicides: DNA Testing of Retained Autopsy Sexual Assault Smears

Archival medical examiner specimens may contain perpetrator DNA evidence useful in unsolved (“cold case”) homicides. The Office of the Chief Medical Examiner (OCME) histology slide archives were searched for sexual assault smears for all 376 female homicides from 1990 to 1999. Of these, the OCME had sexual assault smears on 84 of which 13 slides had sperm. Of these 13, six were still unsolved. DNA profiles were obtained on all six (5 from smears and one from swabs). Combined DNA Index System ( submission resulted in two matches (“hits”) for new suspects. In addition, three suspects were eliminated in two cases. Our review of archival sexual assault smears resulted in DNA profiles that were able to assist in the investigation of four cold case homicide investigations. It may be worthwhile for medical examiner offices to search their archival histology slides for sexual assault smears on previously unsolved cases particularly those prior to the mid‐1990s when DNA testing was less widely available.     

Automation of the Differential Digestion Process of Sexual Assault Evidence,

Sexual assault evidence samples require the use of a specific process known as a differential digestion to separate sperm from nonsperm cells prior to DNA extraction. An automated differential digestion process was developed using a selective degradation technique, which uses DNase I to digest the remaining nonsperm DNA in the sperm fraction. The use of DNase on pristine samples, as well as aged and degraded samples, was assessed to ensure that the quantity and quality of the sperm DNA were not compromised or adversely affected. Samples processed using the selective degradation technique yielded comparable DNA yield and DNA typing data to the conventional differential digestion process. The automated process utilized 96‐well plates for high throughput and incorporated microscope slide preparations for confirmation of sperm. It reduced processing time by about sixfold and was paramount in the elimination of the Oakland Police Department Criminalistics Laboratory's sexual assault kit backlog.     

Persistence of vaginal spermatozoa as assessed by routine cervicovaginal (Pap) smears

Retrospective review of cervicovaginal (Pap) smears of women with known sexual histories affords an opportunity to assess the potential for postcoital sperm recovery for large numbers of individuals. This study reviewed 542 individuals' Pap smears with accompanying sexual histories. Three hundred forty-nine respondents reported at least one act of sexual intercourse during the five days preceding the Pap smear. Unlike a previous report, the current study showed very poor sperm recovery (maximum of 25% during the first postcoital day). The observed results roughly correlate with the incidence of sperm noted on screening of large numbers of routine clinical Pap smears in a private reference laboratory. Routine Pap smears can detect sperm but do not appear to be an ideal method to substantiate recent sexual intercourse.     

STR and Y-STR genotyping of 30–50-year-old semen stains

This study performed the semen discrimination test, short tandem repeat (STR) typing, and Y-chromosome specific-STR (Y-STR) typing on five, 30–50-year-old semen stains. All samples reacted positively with the SM test reagent, and we observed sperm heads in all samples microscopically. The quantity of DNA extracted from the 43- and 50-year-old samples was much lower than from the other samples. STR typing of the 30-, 32-, 32-, 43-, and 50-year-old semen samples detected a maximum of 13, 15, 15, 11, and 6 of 15 loci, respectively, while Y-STR typing detected 16, 16, 16, 10, and 10 of 16 loci. These results suggest that the semen discrimination test and STR and Y-STR typing can detect extremely old semen stains and are useful for forensic practice.     

Laser microdissection separation of pure spermatozoa from epithelial cells for short tandem repeat analysis

Short tandem repeat (STR) analysis is a valuable tool in identifying the source of biological stains, particularly from the investigation of sexual assault crimes. Difficulties in analysis arise primarily in the interpretation of mixed genotypes when cell separation of the sexual assailant's sperm from the victim's cells is incomplete. The forensic community continues to seek improvements in cell separation methods from mixtures for DNA typing. The feasibility of applying laser microdissection (LMD) technology to precisely separate sexual assault cell mixtures by visual inspection coupled with laser dissection was assessed through three experiments. First, various histological stains were evaluated for use with LMD and DNA analysis. Second, different DNA isolation methods were evaluated on LMD-collected cells. Finally, STR analysis was performed on LMD-separated sperm cells from mixtures of semen and female buccal epithelial cells. The results indicated (a) hematoxylin/eosin staining performed best in its ability to differentiate sperm and epithelial cells while exhibiting the least negative effect on further downstream analysis; (b) both QIAamp and Lyse-N-Go methods were useful for recovery of DNA from LMD-collected sperm cells; and (c) LMD separation provided clear STR profiles of the male donor with the absence of any additional alleles from the female donor. This report describes an efficient, low-manipulation LMD method for the efficient separation of spermatozoa from two-donor sperm/epithelial cell mixtures.     

DNase-Ⅰ纯化结合碱性裂解法提取混合斑精子DNA

目的研究脱氧核糖核酸酶I(DNase-I)纯化结合碱性裂解法提取混合斑精子DNA的方法在法医学中的应用。方法收集79份性犯罪案件混合斑检材,分别用DNase-I纯化结合碱性裂解法和差异裂解法提取精子DNA,采用STR荧光标记复合扩增体系进行16个STR基因座分型,并比较检验结果。结果应用DNase-I纯化结合碱性裂解法提取精子DNA,64例检材分型成功;应用差异裂解法提取精子DNA,57例检材分型成功;两种方法比较结果存在显著性差异(P=0.039),DNase-1纯化结合碱性裂解法提取精子DNA的STR分型成功率更高,成本低廉。结论DNase-I纯化结合碱性裂解法提取混合斑精子DNA可提高检验成功率,操作简便,快速,易于自动化,适于法医学个体识别鉴定。     

DNase-I纯化结合碱性裂解法提取混合斑精子DNA

目的研究脱氧核糖核酸酶I(DNase-I)纯化结合碱性裂解法提取混合斑精子DNA的方法在法医学中的应用。方法收集79份性犯罪案件混合斑检材,分别用DNase-I纯化结合碱性裂解法和差异裂解法提取精子DNA,采用STR荧光标记复合扩增体系进行16个STR基因座分型,并比较检验结果。结果应用DNase-I纯化结合碱性裂解法提取精子DNA,64例检材分型成功;应用差异裂解法提取精子DNA,57例检材分型成功;两种方法比较结果存在显著性差异(P=0.039),DNase-1纯化结合碱性裂解法提取精子DNA的STR分型成功率更高,成本低廉。结论DNase-I纯化结合碱性裂解法提取混合斑精子DNA可提高检验成功率,操作简便,快速,易于自动化,适于法医学个体识别鉴定。     

葡萄胎的DNA检验

目的探讨一例强奸案中的葡萄胎样本的DNA检验及结果分析。方法用Identifiler试剂盒对葡萄胎样本和受害人血样进行荧光复合STR基因座扩增并分型。结果该例的葡萄胎在15个STR基因座上分型均为纯合子,应属于单精子受精的完全性葡萄胎。结论葡萄胎类型多种,对应着不同的DNA分型,在实际案件的检验中应引起注意。