人类D19S40基因座在不同人种中的遗传多态性研究

采用ＰＣＲ技术分析中国汉族、德国人、斯洛伐克人和美国黑人群体Ｄ１９Ｓ４００基因座的遗传多态性及世界三大人种之间的差异。四个群体共调查了６２０人,发现了１１个等位基因,观察到４７种基因型。各群体观察杂合度为：０．７８～０．８８,个人识别机率为：０．９３８５０～０．９６６４。四个群体基因型频率分布符合Ｈａｒｄｙ－Ｗｅｉｎｂｅｒｇ平衡（Ｐ＞０．０５）,三大人种（蒙古人种、高加索人种、美国黑人）之间Ｄ１９Ｓ４００基因座等位基因频率分布存在极显著差异（Ｐ＜０．０１）。结果显示Ｄ１９Ｓ４００基因座在群体遗传学研究和法医学个人识别中有较高应用价值     

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（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     

汉族人群五个STR基因座的多态性调查

应用PCR及PAG电泳技术研究了PLA2A、vWA、CYP19、TH和LPL五个基因座的多态性，调查武汉地区汉族无关个体，获得汉族人群的频率分布。五个基因座基因型频率分布与Hardy-Weinberg平衡吻合良好、分别计算基因座的杂合度（H）、个人识别能力（Dp）、非父排除率（PE）和多态性信息总量（PIC）。为法医学应用提供了基础数据。     

D7S817,D18S865位点的群体遗传学研究

目的：调查D7S817、D18S865两个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位点有成都汉族群体中发现9个等位基因,23种基因型,杂合度为0.738,个人识别几率为0.931。在甘肃东乡群体中发现8个等位基因,20种基因型,杂合度为0.752,个人识别几率为0.917。D18S865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,个人识别几率为0.906;在甘肃东乡群体中发现6个等位基因,15种基因型,杂合度为0.814,个人识别几率为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、D18S865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D20S85和D6S477基因座遗传多态性调查

目的对武汉地区280名汉族无关个体的D20S85和D6S477位点遗传多态性进行调查,研究其在法医学检验中的应用价值。方法应用PCR和PAGE技术进行分型检验。结果分别检出10个、9个等位基因,获得各等位基因在该地区汉族人群分布频率,基因型频率分布符合Hardy-Weinberg平衡。两位点的DP值分别为0.9085、0.9127。结论D20S85和D6S477是法医学中重要的遗传标记。     

vWFⅢ基因座扩增片段长度多态性及其法医学应用

研究短串联重复vWⅢ基因应扩增片段长度多态性及其法医学意义。应用STR－PCR分型法对200名中国辽宁汉族无关个体的短串联重复VWFⅢ的多态性进行调查。共检出8个等位基因,频率分布在0．003～0．333,基因长度范围为154～182bP,个人识别能力为0．921,非父排除率为0．581。群体调查证实该基因座符合Hardy－Weinberg平衡,家系调查结果表明其遗传符合孟德尔方式。该基因座可作为人类群体遗传学的一个有价值的多态性标记,可用于法医学个人识别及亲子鉴定。     

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。     

短串联重复位点ACTBP2(SE33)的扩增片段长度多态性研究

应用变性聚丙烯酸胺凝胶电泳（dn－PAGE）结合银染色技术对短串联重复（STR）位点ACTBP2（SE33）的扩增片段长度多态性（Amp－FLPs）进行了研究。在210名无关中国个体中观察到了25个等位基因，等位基因频率分布在0．007～0．093之间。基因型的分布符合Hardy－Weinberg定律，个体识别能力（DP）值为0．99，杂合度（H）为98．7％。七个家系分析的结果表明，该位点的遗传符合孟德尔遗传法则，未观察到变异。对几种常见的法医物证检材的分析表明，该分型系统对DNA降解放为严重的检村适用性强，而且灵敏度高（0．5ng），适合于法医学实际应用。     

中国成都汉族及泰国群体D7S2846基因座的遗传多态性

研究STR基因座D7S2 846的遗传多态性 ,为法科学应用提供基础数据。应用PCR及PAG电泳技术 ,对376名中国成都汉族无关个体及 131名泰国无关个体进行了调查。两群体分别检出 8个和 7个等位基因 ,首次获得该基因座基因在两群体中的频率分布。两群体基因型频率分布均符合Hardy Weinberg平衡。家系调查证实了等位基因的传递遵循孟德尔遗传规律。该基因座在两群体中的个人识别能力 (Dp)分别为 0 85 70、 0 86 0 2 ,杂合度 (H )分别为0 6 915、 0 6 870 ,多态性信息含量 (PIC)分别为 0 6 445、 0 6 5 5 3 ,非父排除率 (PE )分别为 0 415 2、 0 40 85。D7S2 846基因座在法医学个人识别及亲子鉴定中具有较高的实用价值。     

新Y-STR基因座DYS709在汉族人群中的遗传多态性调查

目的筛选新的Y-STR基因座,调查其在汉族人群中的等位基因频率分布,评价其在法医学及其它方面的应用价值。方法在Y染色体基因组DNA中查找候选基因座,在重复顺序两端设计引物,PCR扩增后用银染法显示结果。结果一个重复单位为CTTT的Y-STR基因座DYS709被发现。在102例汉族无关男性个体血样中共检出了7个等位基因。基因多样性为0.7063,个人识别能力(PD)和非父排除率(PE)均为0.7063。结论新筛选到的DYS709具有较高的遗传多态性,在法医学及人类遗传学方面具有应用价值。     

New alleles and mutational events in D12S391 and D8S1132: sequence data from an eastern German population.

To investigate the DNA mutation rate and pattern in the hypervariable short tandem repeat (STR) locus D12S391 and in the locus D8S1132, samples from an eastern German population (Dresden area) were analysed. A duplex PCR was applied, using short amplification products for D12S391 (129-177bp) and a modified reverse primer for D8S1132 (127-182bp). The sequences of some rare and new variant alleles are described. At the locus D12S391, 13 regular and six incomplete alleles with different lengths were found, exhibiting several sequence structures. Two isolated father/child mismatches were observed in a total of 648 meioses. Novel alleles 13.1, 14.1 and 27 were discovered at the locus D8S1132. Three parent/child mismatches were found in a total of 672 meioses.     

6个miniSTR基因座荧光标记复合扩增的遗传多态性

目的评价6个miniSTR基因座在DNA高度降解检材中的法医学应用价值,并调查广东汉族人群6个miniSTR基因座的遗传多态性。方法采用两个复合扩增PCR体系、四色荧光标记及毛细管电泳技术,对D1S1677,D2S441,D4S2364,D10S1248,D14S1434,D22S1045基因座进行基因型检测。结果6个miniSTR基因座均获得了清晰的基因型分型结果,扩增片段均小于120bp,分别检出7、7、5、8、8、7个等位基因和14、11、11、19、12、14种基因型,基因型分布均符合Hardy-Weinberg平衡。6个miniSTR基因座在广东汉族群体的个人识别率和非父排除率分别依次为0.863、0.895、0.792、0.894、0.814、0.904和0.392、0.360、0.353、0.568、0.378、0.513。10例IdentifilerTM试剂盒未能正确分型的高度降解DNA样本,采用6个miniSTR基因座复合扩增体系检测均提高了分型成功率结论6个miniSTR基因座荧光标记复合扩增体系在DNA高度降解检材的检测中具有较高的应用价值,并且在广东地区汉族群体中具有较好的遗传多态性。     

Two loci ‘exclusion’ of true paternity is due to genetic disorder in a child

We describe a paternity case with three genetic incompatibilities between a three-year-old boy and his putative father.STR analysis of 2 out of 25 markers revealed the absence of paternal alleles and presence of two maternal alleles at D2S441 and D2S1338 loci in the child. The rest 23 STR markers served to confirm paternity. In addition, we analyzed Y-STRs and determined the same haplotype in the child and his putative father.With massive parallel sequencing on HID Ion GeneStudio S5 System using Precision ID GlobalFiler NGS STR Panel v2 (Applied Biosystems) we confirmed the presence of two alleles of maternal origin at D2S441, D2S1338 loci and identified two maternal alleles at additional locus D2S1776 located on chromosome 2 in the child.Finally, we confirm paternity. Three loci ‘exclusion’ was due to maternal uniparental disomy of chromosome 2 in the child.     

Allelic structure and distribution of two STR loci, D8S580 and D22S442, in the Japanese population

The allelic frequency and structural characteristics of two STR loci D8S580 and D22S442 were investigated using blood samples from 143 unrelated healthy Japanese individuals. Thirty-eight alleles in D8S580 locus and 13 alleles in D22S442 locus were identified. The discrimination power, heterozygosity, and the polymorphic information content of those loci displayed high values (0.98, 0.88, and 0.87 in D8S580 and 0.97, 0.86 and 0.85 in D22S442), and their frequency distributions met Hardy-Weinberg equilibrium expectations. The allelic pattern of D8S580 was complex and differentiated into three groups (group I: alleles 184-194bp; group II: alleles 203-223, 235, 239, 243, 252 and 255bp; group III: alleles 227-286bp). Most of their alleles contained five categories of repeat units (A: aaaag; B: aaag; C: aagg; D: caag; E: agaa). On the other hand, D22S442 contained only two types of repeat units (A: agga; B: aggg). The present study, hence, proves that both D8S580 and D22S442 are highly polymorphic and represent stable genetic markers applicable to forensic investigations.     

Genetic variation and population genetic data of the short tandem repeat locus D8S320

The locus D8S320 is an STR system first described in 1993 as a simple (AAAG) repeat. Sequencing data revealed that the D8S320 locus is a complex STR system consisting of (AAAG)- and (AAAC)-repeat units. A total of 22 different alleles were found in a population survey of 210 unrelated individuals from the Rhine area with frequencies ranging from <0.01 to 0.198. The population data revealed the existence of variant alleles differing by 1 and 2bp from the consensus allele. Due to the complex repeat structure consisting of three variable regions and one constant region, electrophoresis under denaturing conditions is strongly recommended. The statistical values were calculated to be 0.89 (observed heterozygosity rate), 0.96 (discrimination index) and 0.71 (mean exclusion chance). No deviation from Hardy-Weinberg equilibrium (HWE) was observed.     

Allele frequencies for six miniSTR loci of two ethnic populations in China

Allele frequencies and forensic parameters for the six miniSTR loci D1S1677, D2S441, D4S2364, D10S1248, D14S1434, and D22S1045 were investigated in two ethnic China populations. Allele frequencies for each locus are reported along with nomenclature based on sequence analysis. The polymerase chain reaction (PCR) products contained within the six loci were less than 125 bp in size. All loci showed a moderate degree of polymorphism with observed heterozygosity >0.7 except for D4S2364 and did not show departures from Hardy-Weinberg equilibrium for all two populations. The accumulated powers of discrimination for the six loci were 0.999994, and 0.999996 for Han ethnic, and Korean ethnic minority group, respectively.     

Importance of storing emergency serum samples for uncovering murder with insulin

Using the polymerase chain reaction (PCR), we studied the short tandem repeat (STR) polymorphism observed at the D12S391 locus. In 350 Japanese examined, 14 different alleles ranging from 209 bp to 261 bp were detected. Allele 18 (221 bp) showed the highest frequency at 0.30. Observed and expected values of respective genotypes satisfied the Hardy-Weinberg equilibrium (chi 2 = 24.08, P = 0.24, df = 20). In addition, 18 additional sequence structures (suballeles), were detected in this study. Within the suballeles, sequence variants, in which the initial repeat of (AGAT) was replaced with (AGGT), was found in five samples. It was found that the analysis of single-strand conformation polymorphism (SSCP) before sequence analysis was useful for distinguishing these suballeles.     

3个miniSTR基因座在湖南汉族人群中的遗传多态性

目的建立扩增片段小于120bp,包括D10S1248、D2S441和D1S16773个miniSTR基因座的复合扩增系统,并调查其在湖南汉族人群中的遗传多态性。方法采用不同荧光染料标记引物,通过PCR扩增,利用ABI 310遗传分析仪对186份无关个体血样进行3个miniSTR基因座片段长度分析。结果D10S1248、D2S441和D1S1677 miniSTR基因座均获得了清晰的基因分型结果,经对186名无关个体进行分析,分别检出9、7、7个等位基因和21、19、15种基因型,基因型分布符合Hardy-Weinberg平衡。3个基因座在湖南汉族人群的非父排除率和个体识别力分别为0．465、0．491、0．361和0．886、0．899、0．818。结论建立的3个miniSTR基因座扩增系统在DNA高度降解检材分析中具有较高的应用价值,并且在湖南汉族人群中具有较好的遗传多态性,可应用于个体识别和亲权鉴定。     

Allele frequencies of four VNTR loci in the Chinese population in Hong Kong

DNA specimens isolated from the blood samples of 209 unrelated Chinese persons were characterised with four probes, i.e. MSI (D1S7), YNH24 (D2S44), pH30 (D4S139) and TBQ7 (D10S28), in conjunction with the restriction enzyme HaeIII to establish a population DNA-profile database. Each locus showed a fairly wide distribution of alleles and high heterozygosity. The sizes of alleles ranged between 0.73 and 19.95 kb for the D1S7 locus, between 0.78 and 4.72 kb for the D2S44 locus, between 2.53 and 15.74 kb for the D4S139 locus, and between 0.66 and 6.53 kb for the D10S28 locus. The heterozygosities for the D1S7, D2S44, D4S139 and D10S28 loci were 97.6%, 92.3%, 86.6% and 96.2%, respectively. Allele frequencies were determined using the ‘fixed-bin’ approach. The bins were based on the 30-band pattern of the molecular size marker used in this laboratory (GIBCO BRL 4401SA). The most commonly observed bin frequencies for the four loci ranged from 9% to 19%. When a default minimum bin frequency of 10% and the product rule based upon the principles of Hardy-Weinberg equilibrium were used to calculate the most common genotype frequencies, the values ranged from 2.1% to 6.4%. The minimum composite discriminating potential of the four loci, expressed as the product of the frequency of the most common genotype for each locus, was about 1 in 770 000.