中国壮、蒙、哈萨克族群体30个InDels的遗传多态性

本文对中国地区广西壮族、蒙族、哈萨克族三个健康无关个体进行遗传多态性调查。采用Investigator?DIPplex试剂盒对广西壮族、蒙族、哈萨克族的150名无关个体的30个In Del位点进行复合扩增,统计分析各位点的频率数据、群体遗传学参数。经Bonferroni校正,30个In Del位点不存在连锁不平衡现象;基因型在三个民族的分布均符合Hardy-Weinberg平衡;各位点在蒙族、哈萨克族、广西壮族三个人群中的累积个体识别率分别为0.999 999 999 991 9、0.999 999 999 999 32、0.999 999 999 969,累积非父排除率(CPE)均大于0.98;30个In Dels在蒙族、哈萨克族、广西壮族分别有中2个、1个、6个位点的He0.3。通过Genepop 4.2软件计算Fst值,除D118(0.126)、D64(0.071)外,其余位点均小于0.06。所调查人群具有较高的多态性,能达到较高的个体识别能力,可以作为现有STR检验体系的补充。     

云南壮族人群15个STR基因座遗传多态性

目的对600个云南壮族人群15个常染色体短串联重复(short tandem repeat,STR)序列基因座的遗传多态性进行调查,探讨云南壮族群体遗传学特性及在法医学中的应用价值。方法使用AmpFLSTR^■Identifiler^■Direct PCR扩增试剂盒对此地区600名壮族无关个体血样DNA进行PCR扩增,采用Modified Powerstats软件计算等位基因频率及法医学参数(观察杂合度、期望杂合度、个体识别率、多态信息含量),应用Arlequin 3.5软件检验各基因座Hardy-Weinberg平衡及连锁不平衡,再比较其他群体的遗传距离。结果云南壮族各基因座个体识别能力(DP)值分布在0.7790~0.9744之间,累计个体识别概率(CDP)达到0.99999999999999999156,非父排除率值范围在0.2869~0.7213,云南壮族与贵州布依族、贵州侗族、广西毛南族、广西仫佬族遗传距离比较接近。结论该试剂盒的15个STR基因座在云南壮族人群中具有高度的多态性,可供法医学个体识别和亲权鉴定,亦可为群体遗传学种族迁徙、语言学语言分支区分、社会学风俗探究等提供基础性研究数据。     

67个X-SNP位点的分型检测和连锁不平衡检验

目的 筛选一组在中国汉族人群中具有法医学应用前景的X-SNP位点.方法 根据dbSNP和HapMap两个数据库提供的位点信息和频率数据从X染色体上筛选出67个候选X-SNP位点,采用多重PCR联合基质辅助激光解析电离飞行时间质谱技术检测中国汉族人群428名无关个体,获得67个候选X-SNP位点在中国汉族人群中的频率数据...     

中国北方汉族人群FUT6基因3个SNPs遗传多态性

目的对中国北方汉族人群FUT6基因编码区序列特征及等位基因多态性进行调查。方法测序分析30例中国北方汉族人FUT6基因整个编码区序列并鉴定其单倍型,采用复合PCR与复合限制性内切酶结合的RFLP法分析FUT6基因rs778805（C370T）、G855A及rs61147939（C907G）3个SNPs遗传多态性;应用Haploview4.1软件进行相关统计分析。结果 30例测序样本中共检出8个SNPs和6种单倍型,149例中国北方汉族个体C370T、G855A、C907G基因型分布均符合Hardy-Weinberg平衡。杂合度分别为0.544、0.416、0.510;多态信息含量为0.375、0.372、0.367;个人识别能力为0.600、0.651、0.603;非父排除率为0.187、0.186、0.183。PCR-RFLPs检出13种基因型,单倍型变异度为0.646。Haploview4.1软件分析表明3个SNPs处于连锁不平衡状态。结论中国北方汉族人群FUT6基因编码区序列呈现出高度多态性;C370T、G855A、C907G位点多态性分布良好,但处于连锁不平衡状态。     

甘肃省汉族人群18个STR位点的遗传多态性

目的 调查18个短串联重复序列（Short Tandem Repeat,STR）位点在甘肃省汉族人群中的基因频率分布.方法 采用PCR扩增及毛细管电泳技术对272名个体的18个STR基因座进行分析.结果 共检出202种等位基因,基因频率分布在0.002～0.570之间.18个STR基因型分布均符合Hardy-Weinberg平衡（P＞0.05）,杂合度在0.599～0.893之间,个人识别能力在0.771～0.984,多态信息含量在0.534～0.910,非父排除概率在0.290～0.782.结论 本研究结果可为人类群体遗传学及法医学后续研究提供详实可靠的基础数据.     

浙江宣城地区汉族人群18个STR基因座遗传多态性

目的 调查浙江宣城地区汉族人群5000例无关个体18个STR基因座多态性分布.方法 应用AmpFlSTR Identifiler荧光标记复合扩增系统检测,统计计算群体遗传学参数.结果 18个STR基因座的基因型分布均符合Hardy-Weinberg平衡（P＞0.05）,共检出270个等位基因,频率在0.0001～0.5260之间,共有1289种基因型,在13个基因座上检出共44个微变异等位基因.18个基因座的杂合度观察值（Ho）在0.6238～0.9120之间,杂合度理论值（He）在0.6961～0.9601之间,多态信息含量（PIC）在0.5578～0.9126之间,累计个人识别率（TDP）为0.999 999 999 999 999 999 999 953,三联体累计非父排除概率（CPE）为0.999 999 993487 306,二联体累计非父排除率（CPE＊）为0.985819169.结论 本文调查结果与以往文献报道的不同地区民族的人群等位基因频率资料有一定程度的差异性.上述18个STR基因座适用于宣城地区汉族群体各类案件的法医学个体识别和亲权鉴定.     

高原适应基因中藏族祖先信息位点的研究

目的基于高原适应基因上的SNPs位点筛选出藏族祖先信息位点。方法本研究利用Mass ARRAY?基因分型系统对183份藏族个体在EGLN1和EPAS1基因上的87个SNPs位点进行分型检测,结合千人数据库中26个世界人群2504份样本的分型数据,进行群体间差异分析,筛选出藏族特异性高的位点作为藏族祖先信息位点(Ancestry informative SNPs,AISNPs),并通过群体间遗传结构对筛选出的位点进行分析评估。结果在87个高原适应基因SNPs位点中,有23个位点在藏族群体中的频率分布具有特异性,可作为藏族AISNPs。STRUCTURE聚类分析结果表明:在K=2时,藏族人群具有与其他26个人群不同的遗传主成分。结论本研究筛选出的23个藏族祖先信息位点可合并到现有的祖先推断体系中,进一步增加针对东亚亚人群的分辨率,也可为相关案件提供侦破线索。     

中国北方汉族群体DRD4基因3个位点遗传多态性

目的调查DRD4基因启动子区-1240L/S、-521C/T和第三外显子48bp VNTR 3个位点在中国北方汉族群体的遗传多态性分布,评价其法医学应用价值。方法收集中国北方汉族207例个体血液样本,提取模板DNA,采用聚合酶链反应和等位基因特异性扩增技术,对3个位点进行分型检测,应用Arlequin 3.5软件对分型数据进行统计分析。结果 3个位点分别检出2个(-1240L/S、-521C/T)和6个(48bp VNTR)等位基因,3种和9种基因型,经χ2检验,各位点基因频率分布均符合Hardy-Weinberg平衡(P0.05);DP值均超过0.5;3个位点共检出14种单倍型,其中8种为主要单倍型,DP值为0.940,PE值为0.804;-1240L/S位点与非洲及高加索人群间的差异具有统计学意义(P0.05),而与日本人群以及-521C/T位点与3种群体之间均无显著性差异(P0.05)。结论中国北方汉族人群DRD4基因3个位点多态性分布均较好,在法医学个体识别与亲子鉴定中具有一定的应用价值。     

30个 InDels在中国三个民族的遗传学调查及法医学应用

目的调查Qiagen Investigator@ DIPplex试剂盒30个InDels多态性位点在中国汉族、藏族、维吾尔族人群中的群体遗传学数据,评估其法医学应用价值。方法采集汉、藏、维吾尔族各90名无关个体静脉血,提取DNA。使用3130xL毛细管电泳对该270份样品进行分型,通过统计计算相关的群体遗传学参数。结果实验得到270份样品的分型及基因型频率,30个InDels未明显偏离Hardy-Weinberg平衡及连锁平衡,在汉族、藏族、维吾尔族三个人群中的随机匹配概率分别为1.42×10（-11）、7.19×10（-12）、4.74×10（-13）,累积非父排除率（CPE）均大于0．9951。结论该组插入缺失位点在中国的汉族、藏族、维吾尔族人群中具有较高的多态性,能达到较高的个体识别能力,可以作为现有STR检验体系的补充。     

江苏汉族人群30个插入/缺失位点的遗传多态性

目的调查30个插入/缺失(insertion/deletion,In Del)位点在江苏汉族人群中的遗传学信息,评估Investigator~ DIPplex试剂盒的使用价值,并用于指导江苏汉族人群的法医学分析。方法用Investigator~ DIPplex试剂盒对江苏地区305名汉族健康无关个体进行常染色体In Del位点的分型检测,统计分析30个In Del位点的频率数据及遗传学参数。结果 30个In Del位点在江苏汉族人群中的分布均符合Hardy-Weinberg平衡,有21个In Del位点的最小等位基因频率大于0.3。多态信息含量为0.089~0.375,个体识别率为0.093~0.500,二联体非父排除率为0.047~0.250,三联体非父排除率为0.046~0.219。30个In Del位点经连锁不平衡分析,各位点之间相互独立,累积个体识别率为1-7.369×10-8,二联体累积非父排除率为0.998 933 978,三联体累积非父排除率为0.997 806 392。除HLD118等5个位点外,群体间Fst值均小于0.06,在群体间差异较小。结论 Investigator~ DIPplex试剂盒中所含有的In Del位点可作为补充遗传标记应用于法医物证学相关检案工作。     

Genome‐wide Screen for Individual Identification SNPs (IISNPs) and the Confirmation of Six of Them in Han Chinese with Pyrosequencing Technology*

Abstract: Single nucleotide polymorphisms (SNPs) offer promise to forensic DNA analysts, but it remains uncertain whether a panel of individual identification SNPs can be as informative as the Combined DNA Index System short tandem repeats. Based on the highly accurate and publicly available HapMap SNP database (r21a) and a minor allele frequency cutoff of ≥0.45, we completed a genome‐wide screen through 3,905,819 SNPs with internally modified computer programs and identified 1439 SNPs with high heterozygosity and low F_st values among four populations (Utah Caucasian, Han Chinese, Tokyo Japanese, and Nigerian Yoruba). Using pyrosequencing technology, we studied six loci in a relatively large group of samples to determine whether these loci were as informative as the HapMap data suggest. These SNPs performed as expected in the Han Chinese in terms of heterozygosity and F_st. The 1439 identified SNPs should provide a comprehensive and reliable set of loci for identity and relationship testing.     

Evaluation of 96 SNPs in 14 populations for worldwide individual identification

Great advances have been made recently in searching for individual identification single-nucleotide polymorphisms (IISNPs or IDSNPs). Such SNPs as suggested by SNPforID scientists and by Pakstis et al., are promising, although they were selected from older or smaller databases rather than the most recent database. Here, we describe a new computational strategy for developing IDSNPs based on HapMap. We searched through HapMap r27 for SNPs having minor allele frequencies ≥0.30 in all its 11 populations and found more than 1881 qualified SNPs. We examined 96 of them with 183 DNA samples from three Chinese populations using Illumina arrays. The average allele frequency for these 96 SNPs among the three populations was 0.495/0.505, the average number of identical SNP genotypes shared by two individuals among the 14 populations (three Chinese and 11 HapMap) was 37.9, and the random matching probability for two unrelated Hans to match in all 96 genotypes was 9.793 × 10(-39). Thus, most of these 96 SNPs are universally applicable.     

法医SNP复合检测体系的构建及应用

目的构建48-SNP位点复合检测体系,用于个体识别、性别鉴定、ABO基因分型。方法采集225份无关个体样本(血斑及口腔拭子),18份案例样本(不同组织及体液斑),选择43个常染色体位点、4个ABO基因位点和1个性别鉴定位点,根据单碱基延伸技术通过GenomeLabTMSNPstream基因分型系统进行SNP分型;并检测体系灵敏度、同一个体不同组织同一性及模拟腐败检材。结果 48-SNP体系分型结果与测序结果的一致性为100%,最小DNA检出量为0.25ng,不同组织来源样本检测同一性很好;利用该体系检测225名无关汉族个体,所有位点均符合Hardy-Weinberg平衡,整个系统的随机匹配概率为9.4×10-18,累积非父排除率(CEP)为0.999 788,累积个体识别率大于0.999 999 999 999 999 99。结论本文48-SNP体系能同时进行个体识别、ABO基因分型和性别鉴定,可以作为现有STR检验体系的补充。     

Forensic Identification Using a Multiplex Assay of 47 SNPs*

Abstract: As a powerful alternative to short tandem repeat (STR) profiling, we have developed a novel panel of 47 single nucleotide polymorphisms (SNPs) for DNA profiling and ABO genotyping. We selected 42 of the 47 SNPs from a panel of 86 markers that were previously validated as universal individual identification markers and identified five additional SNPs including one gender marker and four ABO loci. Match probability of the 42 validated SNPs was found to be 9.5 × 10^?18 in Han Chinese. SNP analysis correctly assessed a panel of historical cases, including both paternity identifications in trios and individual identifications. In addition, while STR profiling of degraded DNA provided information for 11 loci of 16 potential markers with low peak intensities, SNPstream^® genotyping was sufficient to identify all 47 SNPs. In summary, SNP analysis is equally effective as STR profiling, but appears more suited for individual identification than STR profiling in cases where DNA may be degraded.     

应用Ion Torrent PGM~(TM)平台检测中国汉族124个身份鉴定SNPs

目的应用Ion Torrent PGM~(TM)测序平台检测中国汉族群体124个身份鉴定SNPs(individual identification SNPs,IISNP)的多态性信息。方法采用Ion Ampliseq~(TM)Library试剂盒对中国汉族130个无关个体样本及2个家系共8个个体的124个SNPs(90个常染色体SNPs和34个Y染色体SNPs)进行复合扩增,在Ion Torrent PGM~(TM)测序平台上检测。结果中国汉族130个无关个体应得14 148个SNP分型,其中软件给出分型结果14 086个,正确14 085个(99.992 9%),分型偏倚1例(0.007 1%)。软件未报SNP分型62例,需人工校正分析。在90个常染色体SNPs中,MP值最高为0.817 3(rs740910),最低为0.348 0(rs2831700),CMP为6.8984×10~(-34);DP值最高为0.652 0(rs1355366),最低为0.182 7(rs727811),CDP为0.999 999 999 999 999 999 999999 999 999 999 310 2,高于22个STRs的CDP;PE值最高为0.278 1(rs1058083),最低为0.007 3(rs1024116),CPE为0.999 999 616 7,低于22个STRs的CPE。在34个Y-SNPs中,72个中国汉族男性无关个体共观察到8种单倍型。家系样本分型结果未发现突变,均符合遗传规律。结论 124个身份鉴定SNPs在中国汉族群体中具有良好的遗传多态性,是理想的个体识别遗传标记。Ion Torrent PGM~(TM)平台在法庭科学领域有较好的应用价值。     

An STR forensic typing system for genetic individualization of domestic cat (Felis catus) samples

A forensic genotyping panel of 11 tetranucleotide STR loci from the domestic cat was characterized and evaluated for genetic individualization of cat tissues. We first examined 49 candidate STR loci and their frequency assessment in domestic cat populations. The STR loci (3-4 base pair repeat motifs), mapped in the cat genome relative to 579 coding loci and 255 STR loci, are well distributed across the 18 feline autosomes. All loci exhibit Mendelian inheritance in a multi-generation pedigree. Eleven loci that were unlinked and were highly heterozygous in cat breeds were selected for a forensic panel. Heterozygosity values obtained for the independent loci, ranged from 0.60-0.82, while the average cat breed heterozygosity obtained for the 11 locus panel was 0.71 (range of 0.57-0.83). A small sample set of outbred domestic cats displayed a heterozygosity of 0.86 for the 11 locus panel. The power of discrimination of the panel is moderate to high in the cat breeds examined, with an average P(m) of 3.7E-06. The panel shows good potential for genetic individualization within outbred domestic cats with a P(m) of 5.31E-08. A multiplex protocol, designed for the co-amplification of the 11 loci and a gender-identifying locus, is species specific and robust, generating a product profile with as little as 0.125 nanograms of genomic DNA.