D8S384基因座的遗传多态性及其在法医物证检验中的应用

揭示人类自然群体中D8S384基因座的基因型频率,评估D8S384基因座在法医物证中的应用价值,以及建立D8S384基因座的分型方法。用不同基因型PCR产物混合的方法,制备了D8S384等位基因分型标准物,并按照国际法医血液遗传学会DNA委员会推荐的原则命名了等位基因。采用PCR扩增、电泳分析、银染显色的方法,调查了世界3大人种11个群体1103名个体的D8S384基因型。D8S384基因座共有8个等位基因,群体内基因型分布符合Hardy－Weinberg平衡,群体间基因型构成有显著性差异。利用群体数据估计了D8S384基因座的法医学理论应用价值,计算得出D8S384基因座的期望杂合度为0．704±0．014,个人识别机率为0．864。D8S384基因座是一个较好的法医学STR遗传标记。     

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。     

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。     

Further characterization and population data for the pentanucleotide STR polymorphism D10S2325.

Pentanucleotide tandem repeat markers are interesting for forensic sciences, because they may present less stutter on the electrophoretic pattern. We focused on the analysis of the DNA sequence for each allele at the pentanucleotide STR locus D10S2325 in order to understand their structures in the human genome and to construct human allelic ladder, which is necessary for forensic DNA typing. In order to evaluate the forensic applicability of D10S2325 and to construct a preliminary database, the genotype distributions and allele frequencies in three major ethnic groups were investigated. The population samples included Caucasians (Germans), Africans (African Americans), and Asians (Chinese). A total of 520 samples from unrelated individuals was analyzed by Amp-FLP. An example of each allele and new alleles were sequenced. Allele determination was carried out by comparison with a sequenced human allelic ladder made in-house. This pentanucleotide STR provided easily interpretable results. A total of 15 alleles was found in our population samples. Three new alleles were observed and named as alleles 19 and 21 based on the number of repeat motifs, while allele 19 can be divided further into two alleles, 19a and 19 according to analysis of the sequence. No evidence of deviation from Hardy-Weinberg equilibrium was observed. In 64 confirmed father/mother/child triplets no mutation event was observed. Using a maximum likelihood method, the mutation rate was indirectly estimated as 2.5 x 10(-5). These results suggest that D10S2325 is a useful marker for forensic casework and paternity analysis.     

中国成都地区汉族群体5个STR基因座的遗传多态性

采用PCR扩增,聚丙烯酰胺凝胶电泳分析技术,调查中国成都汉族群体DIS1656、D851179、D9S302、D185535及D195253等5个STR基因座的等位基因频率分布。D1S1656检出11个等位基因,35种基因型;DSS1179检出9个等位基因,32种基因型;D95302检出12个等位基因,50种基因型;D185535检出7个等位基因,20种基因型;D195253检出8个等位基因,28种基因型。5个STR基因座基因型频率分布符合Hardy-weinberg平衡（P＞0．05）。个人识别机率（DP）为0．92～0．98。分析了二代3口之家的遗传模式,证明5个STR基因座均符合孟德尔遗传规律。5个STR基因座PCR扩增采用同一条件,方法简单、快速、灵敏、重复性好,可用于法科学亲子鉴定和个人识别。     

Allelic frequencies for the HLA-DQA1, D1S80, HUMTHO1, HUMTPOX, HUMCSF1PO and HUMVWA loci in Cantabria (middle north Spain)

Allele frequencies for six DNA polymorphisms have been studied in a population sample from Cantabria (middle north Spain) using the polymerase chain reaction. The HLA-DQA1 locus was analyzed by the reverse dot-blot technique and the other five by polyacrylamide gel electrophoresis followed by silver staining. Six alleles were found for HLA-DQA1. 15 alleles for D1S80, 6 alleles for HUMTHO1 and HUMCSF1PO, 7 for HUMTPOX and 8 alleles for HUMVWA. The 21 repeat allele in HUMVWA had not previously been reported in a Spanish population. The genotype distributions met Hardy-Weinberg expectations for all the systems and some statistical parameters of forensic interest were calculated. Comparisons with other populations revealed significant differences for HLA-DQA1, HUMVWA and HUMTHO1, with interracial differences being more pronounced than between Spanish populations. The HUMVWA system showed the highest forensic efficiency of the six polymorphisms studied.     

Jordanian population data on the PCR-based loci: LDLR, GYPA, HBGG, D7S8 and GC.

Genotype and allele frequency distributions for PM polymerase chain reaction (PCR)-based genetic markers were determined in a Jordanian sample population. Results were obtained using the AmpliType PM PCR Amplification and typing kit. All loci were in agreement with the Hardy-Weinberg equilibrium expectations. The predominant alleles for LDLR, GYPA, HBGG, D7S8 and GC loci were B, A, B, A and C respectively. No statistically significant variation was detected in allele frequencies of these loci in Jordanians compared to that in Israeli Arab, U.S Caucasian and Japanese populations. Data presented here can be used to estimate the frequency of a specific DNA profile in the Jordanian population for forensic analyses and paternity testing.     

PCR-based DNA analysis of vWA31A,TH01, F13A01, FESFPS,TPOX and CSF1PO

The allele and genotype frequencies of 6 tetranucleotide STR loci were investigated in a sample of 132 unrelated individuals from Chinese Han population. The PCR products were analyzed on 6% denaturing PAGE and detected using fluorescently labeled primers in an automated 377 sequencer(PE). All loci meet Hardy-Weinberg equilibrium. There was no random association of alleles among the 6 loci. The allele frequencies were compared with other population databases. Except locus vWA31A, the observed heterozygosity at other 5 loci was significantly lower than that reported in Caucasian and Black population studies. The calculated DP = 0.99999, PE = 0.9708, pM = 1.059 x 10(-5). The allelic frequency data can be used in forensic identification and paternity testing.     

D19S400基因座在中国3个汉族群体与日本人中的遗传多态性研究

为了解中国广州、吉林、成都三个地区汉族和日本人群体D19540O基因座基因频率分布，并获得中国三个汉族群体和日本群体D19M00基因座的群体遗传数据，比较它们之间的遗传学差异，探究在法医学应用中的意义。应用PCR扩增技术，聚丙烯酸胺凝胶垂直板电泳对D19S400基因座分型。在四个群体469个个体中共检出11个等位基因，45种基因型，基因型频率分布均符合Hardy-Weinberg平衡。各群体的观察杂合度为0.75～0.84，非父排除概率为0.6057～0.6582，个人识别机率为0．9301～0．9480。四个群体之间基因频率分布无显著性差异（P＞0．05）。结果表明，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。     

Genotyping of five short tandem repeat loci via triplex and duplex PCR

We have developed a triplex PCR method for D3S1359, HumTH01 and HumTPO tetranucleotide loci and a duplex PCR method for HumFES/FPS and HumvWA31A tetranucleotide loci using high resolution polyacrylamide gel electrophoresis and silver staining. The methods were evaluated for paternity testing and individual identification and allele frequencies at these loci are reported for 189–3387 unrelated individuals in the Finnish population. The D3S1359 locus, especially, was found to be a highly informative locus. Seventeen alleles were found in the D3S1359 locus with a highest observed allele frequency of 0.199, a high exclusion power (PE) in paternity testing (0.78) and a high observed heterozygosity (0.89). The combined PE for these five loci was 0.99.     

D6S2418在中国（汉族）、泰国和德国人群中的遗传多态性

目的获得D6S2418基因座的群体遗传学数据,分析其基因频率在不同群体间的分布情况是否存在差异,并分析其在法医学中的应用价值。方法采用PCR、非变性聚丙烯酰胺凝胶电泳(PAGE)及银染技术分析中国成都地区汉族、泰国曼谷地区泰国人群以及德国Maint地区德国人群中D6S2418基因座的遗传多态性,获得三个群体D6S2418基因座的群体遗传学数据。结果从300份分别采自成都地区汉族、泰国曼谷地区泰国人群以及德国Maint地区德国人群三个群体的无血缘关系个体的静脉血,共发现9个等位基因,观测到31种基因型。观测杂和度为64%～81%,个人识别机率为84.2%～93.5%,经统计学检验,基因型的频率分布符合Hardy-Weinberg平衡定律。等位基因频率的分布在三个群体间有显著差异。结论D6S2418基因座的个人识别能力高,在法医学个人识别和亲子鉴定应用中有较高价值。     

成都地区汉族人群D2S441位点的遗传多态性研究

为研究 STR位点 D2S441的遗传多态性,为法医学应用提供基础数据,应用 PCR及 PAG电泳技术对 260名成都地区汉族无关个体进行了调查,共检出 9个等位基因及 26种基因型,首次获得汉族群体频率分布 ,其等位基因片段大小范围为 131～ 155bp。该位点基因型频率分布符合 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。其个人识别能力（ Dp）、杂合度（ H）、多态性信息含量（ PIC）和非父排除率（ PE）分别为 0.9084、 0.7885、 0.7390和 0.5778,表明该位点在法医学个人识别及亲子鉴定中具有较高的实用价值。     

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.     

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。     

湖南汉族人群21个STR基因座的遗传多态性(英文)

目的调查湖南地区汉族人群21个STR基因座(D3S1358、D13S317、D7S820、D16S539、Penta E、D2S441、TPOX、TH01、D2S1338、CSF1PO、Penta D、D10S1248、D19S433、v WA、D21S11、D18S51、D6S1043、D8S1179、D5S818、D12S391和FGA)的遗传多态性。方法共采集560例湖南汉族健康无关个体血液样本,使用Chelex-100法提取DNA,应用AGCU EX22试剂盒及9700 PCR扩增仪进行复合扩增,扩增产物使用310遗传分析仪进行分离分析。结果共发现248个等位基因,等位基因频率分布在0.001~0.518。除Penta E(P=0.023)外,其余基因座的基因型分布均符合Hardy-Weinberg平衡。21个基因座的累积个人识别率、累积非父排除率、累积匹配率分别为0.999 999 999 999 999 999 999 999 8、0.999 999 998和1.36×10-25。结论 21个STR基因座在湖南汉族人群中呈高度多态性。本研究可为法医学个人识别及亲子鉴定提供有价值的数据及理论基础。     

Population studies and validation of paternity determinations by six microsatellite loci

A single locus system of 6 microsatellite markers was evaluated for paternity testing. A nonradioactive method based on peroxidase labeling of a DNA probe was used to estimate the allele frequency of markers D1S216, D3S1217, D7S480, D9S157, D13S153, and D16S422 by genotyping 1134-1698 chromosomes. The number of detected alleles were 22, 15, 23, 10, 16, and 19, respectively, and the allele frequency varied from 0.001 to 0.317. The genotype of 87 families, consisting of mother, father, and child was determined. The probability that a random individual will give a positive paternity was evaluated. We conclude that the markers can be reliably typed and give sufficient and reliable information for paternity testing.     

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     

壮族人群3个STR基因座基因频率分布及其法医学应用

研究3个STR基因座（D21S11、HumFGA、D19S253）在广西壮族人群中的基因频率分布及其在实际检案中的应用价值。以自制等位基因Ladder样品作为标准对照,用PCR结合PAGE技术对3个STR基因座的扩增产物进行分型。结果显示：D21S11基因座有14个等位基因,有44个基因型;HumFGA基因座有15个等位基因,40个基因型;D195253基因座有9个等位基因,23个基因型。经检验,3个STR基因座基因型分布均符合Hardy－Weinberg平衡,累计个体识别力（DP）为0．9995。3个STR基因座在壮族人群属高识别力遗传标记系统,在法医学个体识别及亲权鉴定方面有重要价值。     

浙江汉族人群21个非CODIS系统STR基因座的遗传多态性调查

目的调查21个非CODIS系统STR在浙江汉族人群的遗传多态性,为其法医学应用提供基础数据。方法应用AGCU21＋1荧光标记复合扩增系统,对浙江汉族481名无关个体进行21个STR基因座的复合扩增,用ABl3130XL型基因分析仪对扩增产物进行检测,并统计其STR基因座的群体遗传学参数。结果获得21个STR基因座的等位基因频率分布,分别检出8、11、9、10、8、9、6、5、13、9、6、15、8、7、8、9、8、9、9、16、7个等位基因,并分别获得21个STR基因座的H、He、DP、PM及EP等法医遗传学参数。结论21个STR基因座具有较强个体识别能力,可应用于法庭科学中的个体识别与亲权鉴定。