Unusual FGA and D19S433 off-ladder alleles and other allelic variants at the STR loci D8S1132, vWA, D18S51 and ACTBP2 (SE33)

A very short FGA allele *14 and a long D19S433 allele *19.2 were detected and sequenced, as well as the new D8S1132 alleles *12.1, *14 and *15.1. Further new sequence data (vWA allele *18.3, D18S51 allele *11.2, SE33 alleles *24.2, *32, *34 and *37, including the rare variant allele *13.2) are described.     

武汉汉族人群短串联重复序列D5S818， D7S820位点多态性研究

目的研究Ｄ５Ｓ８１８,Ｄ７Ｓ８２０的多态性及法医学应用价值。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技术对武汉地区汉族２３２例无关个体作Ｄ５Ｓ８１８,Ｄ７Ｓ８２０位点分型调查。结果Ｄ５Ｓ８１８和Ｄ７Ｓ８２０位点分别检出８个和６个等位基因,获汉族人群基因频率分布。二位点基因型频率分布符合ＨａｒｄｙＷｅｉｎｂｅｒｇ平衡。位点杂合度分别为０８１２１和０７９３４,个人识别能力分别为０９４１６和０９２５５,非父排除率分别为０５８４２和０５８１６。结论Ｄ５Ｓ８１８和Ｄ７Ｓ８２０ＳＴＲ位点均是高杂合度、高鉴别能力的遗传标记系统,在法医学个人识别和亲子鉴定中有较高实用价值     

D7S817,D18S865位点的群体遗传学研究

目的　调查D7S817、D18S86 5两个STR位点的遗传多态性 ,获得群体遗传学基本数据。　方法　采用PCR和PAG垂直电泳技术、银染显色方法。结果　D7S817位点在成都汉族群体中发现 9个等位基因 ,2 3种基因型 ,杂合度为 0 .738,个人识别机率为 0 .931。在甘肃东乡族群体中发现 8个等位基因 ,2 0种基因型 ,杂合度为 0 .75 2 ,个人识别机率为 0 .917。D18S86 5位点在成都汉族群体中发现 7个等位基因 ,17种基因型 ,杂合度为 0 .72 ,个人识别机率为 0 .90 6 ;在甘肃东乡族群体中发现 6个等位基因 ,15种基因型 ,杂合度为 0 .814,个人识别机率为 0 .898。基因型频率分布符合Hardy -Weinberg平衡定律。等位基因频率的分布在 2个群体之间无显著性差异。　结论　D7S817、D18S86 5位点的扩增效率高 ,重复性好 ,个人识别能力强 ,在法医学个人识别和亲子鉴定应用中有较高的价值。     

武汉汉族人群STR基因座D3S1358、D13S317、D12S391的多态性研究

目的　调查中国武汉地区汉族人群STR基因座—D3S1 358、D1 3S31 7、D1 2S391基因频率分布和群体遗传数据。方法　从 2 0 8个汉族无关个体收集血液标本 ,应用PCR技术及聚丙烯酰胺凝胶垂直板电泳对D3S1 358、D1 3S31 7和D1 2S391基因座分型。结果　D3S1 358检出 7个等位基因和 4 1个基因型。三基因座基因型分布符合Hardy-Weinberg平衡。观察 2 31次减数分裂均未发现突变基因。另外 ,调查结果计算显示D3S1 358、D1 3S31 7和D1 2S391基因座的杂合度 (H)分别为 0 70 98、 0 80 56和 0 84 0 0 ;三个人识别能力 (DP)分别为 0 851 6、 0 9332和 0 952 3;非父排除率 ( pE)分别为 0 4 463、 0 60 1 6和 0 681 8。结论　D3S1 358、D1 3S31 7和D1 2S391基因座在群体遗传学研究和法医学亲子鉴定及个人识别中具有较高实用价值     

D6S1043和D12S391基因座在亲权鉴定中的应用

目的研究D6S1043和D12S391基因座在亲权关系鉴定案件中的应用价值。方法应用荧光标记复合扩增系统对日常检案中所收集的192名汉族无关个体血样DNA进行PCR扩增,用ABI3100-Avant遗传分析仪对扩增产物进行毛细管电泳,用GeneMapperv3.2软件进行基因分型,统计分析D6S1043和D12S391基因座的多态信息。结果在D6S1043和D12S391基因座分别发现12个等位基因,它们在中国汉族人群中的个体识别能力分别为0.9656和0.9510,二联体非父排除率分别为0.573和0.510,三联体非父排除率分别为0.731和0.679。结论D6S1043和D12S391基因座具有高度多态性,在亲权鉴定中具有重要应用价值。     

Additional variability at the D12S391 STR locus in an Austrian population sample: sequencing data and allele distribution

The highly polymorphic STR locus D12S391 was investigated in an Austrian population sample (N=150) by PCR-amplification, comparative detection on native and denaturing polyacrylamide gels and solid phase single stranded sequencing of three size variant alleles and several additional alleles. A total of 15 alleles, distinguishable by size under denaturing conditions, could be detected. No deviations from Hardy-Weinberg equilibrium were observed in the population investigated (P=0.52). Sequencing of size variants designated 17.3 and 18.3 showed an incomplete (GAT) repeat unit at position two of the tandem region. Additional new sequence variants due to varying compositions of the number of (AGAT) and (AGAC) repeats could be identified. Due to distinct electrophoretical mobilities of alleles of the same size but different sequence structures, denaturing detection conditions should be employed when the aim is standardization.     

短串联重复序列D3S1359基因座的多态性

应用PCR、聚丙烯酰胺凝胶垂直电泳及银染技术对STRD3S1359基因座进行多态性调查,首次获得中国汉族人群的基因频率分布数据。检出19个等位基因,59个基因型,基因型频率分布符合Hardy－Weinberg平衡。观察352次减数分裂未发现突变基因,DP值为0．9380,三联体PE值0．6311,二联体PE值0．4568,PIC值0．7824。证实D3S1359是一个高信息含量的STR标记系统。     

中国武汉汉族群体STR D13S325基因座的多态性研究

应用PCR ,聚丙稀酰胺凝胶垂直电泳及银染技术对 2 16例中国汉族人群STRD13S32 5基因座进行了多态性研究 ,首次获得了中国武汉汉族人群基因频率分布资料。检出 11个等位基因 ,30个基因型 ,基因频率分布符合Hardy Weinberg平衡。DP值为 0 92 92 ,三联体PE值 0 6 137,二联体PE值为 0 4370 ,PIC值为 0 775 5。观察 2 0 0次减数分裂未发现突变基因。研究证实D13S32 5是一个高多态性STR基因座标记系统。     

D3S1754、D18S535基因座在中国北方汉族的多态性分析

目的 了解D3S1754、D18S535基因座多态性在中国北方人群中的分布特点及其应用价值。方法 使用PCR、聚丙烯酰胺垂直板电泳及银染的方法。结果D3S1754基因座检出9个等位基因（n=184）,D185535基因座检出8个等位基因（n=201）,两个位点的等位基因频率在群体中的分布符合Hardy-Weinberg平衡（P>0.05）,它们的杂合率（He）分别为0.706和0.807,个人识别机率（DP）分别是0.859和0.934,非父排除率（EPP）分别为0.464和0.629。结论 D3S1754、D18S535两个遗传标记的个人识别率高、非父排除能力较强且能稳定遗传,具有较高的应用价值。     

武汉汉族人群短串联重复序列D5S818,D7S820位点多态性研究

目的 研究Ｄ５Ｓ８１８,Ｄ７Ｓ８２０的多态性及法医学应用价值。方法 应用聚合酶链反应,聚丙烯酰胺凝胶电泳分离及银染是带技术对地区汉族２３２例无关人体作Ｄ５Ｓ８１８,Ｄ７Ｓ８２０位点分型调查。结果 Ｄ５Ｓ８１８和Ｄ７Ｓ８２０位点分别检了８个和６个等位基因,获汉族人群基因频率分布。二位点基因型频率分布符合Ｈａｒｄｙ Ｗｅｉｎｂｅｒｇ平衡。     

成都汉族、甘肃东乡族群体D10S1432和D10S1213 位点的遗传多态性分析

目的：本研究的目的是了解人类基因组中D10S1432及D10S1213两个STR位点在成都汉族和甘肃东乡族群体中的遗传多态性分布及两个群体之间的关系。方法：采用PCR，聚丙烯酰胺凝胶电泳及银染技术，共调查 209例样本，结果L在D10S1432位点上观察到5个等位基因，15种基因型，在D10S1213位点上观察到9个等位基因，31例基因型。两位点的基因型频率在调查的两个嫩体中的分布符合Hardy-Weinberg平衡定律（P＞0．05）。经统计，D10S1432在这两个群体中的杂合度为0．664和0．737。个人识别几率为0．827和0．820。D10S1213的杂合度为0．664和0．657，个人识别几率为0．836和0．882。结论结果表明，D10S1432和D10S1213两个位点在法医学个人识别和亲子鉴定中有较高应用价值。     

Developmental validation of reduced-size STR Miniplex primer sets

This paper describes a developmental validation study of three Miniplex sets covering 12 of the 13 CODIS loci. As these new sets will be used for the analysis of degraded and low level DNA, the validation studies were performed using 100-125 pg of DNA, the lowest input level at which peak balance, peak intensity, and allele consistency were stable. To demonstrate the applicability of the Miniplex sets to forensic casework, these validation studies were completed in accordance with the Scientific Working Group on DNA Analysis Methods (SWGDAM). A range of tests were performed including studies of concordance with standard multiplex kits, sensitivity and reproducibility, and PCR amplification conditions. Additionally, studies of mixtures, nonhuman and environmentally degraded DNA, and simulated forensic samples were performed. Our results demonstrate that Miniplex STR amplification procedures are a robust and sensitive tool for the analysis of degraded DNA.     

D16S539 microvariant or D2S1338 off-ladder allele? A case report about a range overlapping between two loci

All forensic laboratories routinely use commercial kits and softwares for automated typing; in rare cases genotyping misinterpretations or mislabellings occur. This study refers to the investigation on a D2S1338 off-ladder allele mislabelling observed in DNA profile of murdered woman.The Identifiler^® revealed heterozygosity in the range of D16S539, with a presumptive microvariant allele “14.2”, based on assigned size, while PowerPlex^®16 resulted in a homozygosity of allele “11”. Singleplex amplification of D16S539 locus confirmed homozigosity. D2S1338 locus, the closest to D16S1338 in Identifiler^®, genotyped as homozigote “19”, was singleplex amplified. The off-ladder peak was gel-isolated, sequenced and designed as a rare “11” allele variant [(TGCC)6(TTCC)5]. Genotype was finally designed as D16S539 “11,11” and D2S1338 “11,19”.To avoid genotyping misinterpretations or mislabelling, ambiguous genotypes should be established by two commercial kits at least. Furthermore, off ladder alleles as well as allele microvariants should be assigned by direct sequencing. This issue should be considered in Criminal DNA database requirements, that is still under debate in Italy.     

引物结合区点突变致D18S51等位基因丢失

目的确定D18S51基因座是否存在等位基因缺失及其原因。方法应用多个STR试剂盒检测检材以确定D18S51基因座的等位基因缺失情况;重新设计引物对所测D18S51基因座进行单独扩增,并对缺失的等位基因进行测序。结果该案例被检个体的D18S51侧翼序列引物结合区发生突变,致等位基因丢失。结论亲权鉴定时出现不符合孟德尔遗传规律现象,应使用多个试剂盒检测验证以避免父权误判。     

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。     

Two examples of null alleles at the D19S433 locus due to the same 4 bp deletion in the presumptive primer binding site of the AmpFlSTR Identifiler kit

Two cases of allelic loss at the D19S433 locus after multiplex PCR with the AmpFlSTR Identifiler kit (Applied Biosystems) are described. In both cases the failure of PCR resulted in genetic inconsistencies due to opposite homozygosity. After singleplex PCR with published primers additional alleles were observed and Mendelian inheritance was restored. These PCR products were sequenced and in both cases the same 4 bp deletion near the 3′ end of the repeat region was detected in two alleles of different length. The frequency of these null alleles (two events in 1026 allelic transfers) amounts to 0.0019 (95% confidence limits: 0.0002-0.0070).     

Further allelic variation at the STR-loci ACTBP2 (SE33), D3S1358, D8S1132, D18S51 and D21S11

ACTBP2 (SE33), D3S1358, D8S1132, D18S51 and D21S11 are frequently used STR-loci in the forensic field. This study reports sequence data of further new or rare alleles at these loci, varying in length or in sequence, which were detected in course of investigations for various purposes.     

中国5个群体D20S85基因座的遗传多态性

目的 了解中国5个群体D20S85基因座的群体遗传学数据,比较它们之间的遗传学差异,探讨其在法医学应用中的意义。方法 分别收集5个群体622名无关个体的血样,Chelex-100快速抽提法或饱和酚/氯仿法抽提DNA;扩增后经PAGE垂直板电泳、银染,进行D20S85基因座分型。结果 在5个群体622名无关个体中,共检出9个等位基因,并首次在广东汉族和广西壮族群体中检出等位基因14;每个群体基因频率大于0.05的均为6个,D20S85*6为最常见等位基因。5个群体共观察到35种基因型,群体内基因型频率分布均符合Hardy-Weinberg氏平衡,各群体间基因型构成比无显著性差异。观察140次减数分裂未发现突变。各群体的期望杂合度为0.7720～0.7912;非父排除率,在三联体为0.7538～0.7594,二联体为0.3988～0.4297;个人识别率为0.9175～0,9272;多态信息含量为0.7442～0.7656。应用于亲子鉴定和个人识别案例,效果满意。结论 D20S85基因座是法医学应用价值较高的遗传标记系统。