21个非CODIS STR基因座的遗传多态性

目的 调查华东汉族人群21个非CODIS STR基因座的遗传多态性并评价其法医学应用价值.方法 用AGCU 21+1 STR试剂盒,对华东地区汉族225个无关个体的21个非CODIS STR基因座进行扩增,用3130遗传分析仪检测扩增产物,GeneMapper ID v3.2软件进行分型,采用PowerStats v1...     

A new strategy for the discrimination of mitochondrial DNA haplogroups in Han population

Mitochondrial DNA (mtDNA) haplogroup discrimination is interesting not only for phylogenetic and clinical but also for forensic studies. We discriminated the mtDNA haplogroups of 570 healthy unrelated Han people from Zhejiang Province, Southeast China, by comprehensive analysis mutations of the hypervariable segments-I sequence and diagnostic polymorphisms in mtDNA coding region using real-time polymerase chain reaction (RT-PCR), which was compared with the widely used PCR and restriction fragment length polymorphism (PCR-RFLP) method. The results showed that in superhaplogroup M, haplogroup D was the most common haplotype within this assay to 24.6%, and in the other superhaplotype N, haplogroup B and F were the most common groups. Samples re-identified by PCR-RFLP showed the consistent results that were got with RT-PCR. In conclusion, the RT-PCR strategy appears to be an accurate, reproducible, and sensitive technique for the discrimination of mtDNA haplogroups, especially for mass screenings quickly and economically.     

Long Allele Designations at D2S1338 and D19S433 Loci as Influenced by Various Multiplex STR Kits

European forensic laboratories are replacing the STR multiplex kits with the new generation 16/17 STR kits. This study examines the influence of the new generation kits and the new Applied Biosystems 3500xL Genetic Analyzer on the designation of long D2S1338 and D19S433 off‐ladder alleles. Different allele calls were obtained using the new NGM? (Applied Biosystems) and PowerPlex^® ESI? (Promega) kits compared with AmpF?STR^® SGM Plus? kit (Applied Biosystems). Sequence analysis was used to determine accurate allele designation. The new multiplex kits and the 3500xL Genetic Analyzer improved accuracy of long allele designations. DNA databases worldwide include countless profiles obtained by previous kits. Discrepancies between the new and former technologies may cause failure to detect hits. Discordance is expected due to primer sequence differences between various kits. An additional discordance, occurring in long alleles, independent of primer sequence is reported in this study.     

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.     

2个miniSTR等位基因分型标准物的制备

目的采用分子克隆技术制备miniSTR D3S4529和D12ATA63基因座等位基因分型标准物,并评价其应用价值。方法用荧光引物对835份无关个体血卡样本进行扩增并分型检测,筛选2个基因座的等位基因片段,用分子克隆方法制备等位基因分型标准物,并对中国汉族群体进行遗传学调查。结果根据筛选出的等位基因片段制备出分型标准物,各等位基因峰形尖锐,无双肩峰,峰高基本一致,荧光值在4 000 RFU左右。中国汉族人群D3S4529、D12ATA63基因座分别检出7个、11个等位基因,杂合度分别为0.752、0.723,多态信息含量均为0.71。结论通过分子克隆法制备的miniSTR D3S4529和D12ATA63等位基因分型标准物,在法医学研究中具有较高的应用价值。     

16个线粒体DNASNP复合检测体系的建立

目的建立一种复合检测线粒体DNA（mtDNA）单核苷酸多态性（SNP）分型的方法。方法通过设计等位基因特异性引物并结合毛细管电泳分型技术平台,建立包含16个mtDNASNP位点的复合扩增检测体系。对50名汉族无关个体血样进行mtDNASNP检测,并通过直接测序法对其分型结果进行验证。结果50个样本经本检测体系复合扩增后,均得到清晰的SNP分型结果,当模板量在0．5～10pg时能得到较理想的分型图谱。样本的复合检测结果与直接测序法结果完全一致。结论建立的复合检测体系检测mtDNASNP方法灵敏度高、操作简便、分型准确,为针对mtDNA进行简便、有效的中高通量多态性分型提供了一种新方法。     

Genetic Polymorphism of Nine Non-CODIS STR Loci in Hu-nan Province-based Chinese Han Population

Objective To determine the allelic frequency distribution and genetic parameters of nine non-CODIS DNA index systems of the short tandem repeat （STR） loci （D2S1772, D6S1043, D7S3048, D8Sl132, DllS2368, D12S391, D13S325, D18S1364, and GATA198B05）. Methods A total of 353 blood samples were collected, extracted, amplified, and analyzed from unrelated healthy individuals of Han na- tionality in Hunan Province, China. Results One hundred and fourteen alleles were observed in the pop- ulation with corresponding allelic frequencies ranged from 0.001 0 to 0.323 0. For all the nine non-CODIS STR loci, the observed genotypic data showed no significant deviations from the Hardy-Weinberg equi- librium. The Ho, He, PIC, DP, and PE of the studied non-CODIS STR loci ranged from 0.108 0 to 0.1950, 0.805 0 to 0.8920, 0.7700 to 0.8600, 0.9250 to 0.9660 and 0.6070 to 0.7800, respectively. Conclusion Nine non-CODIS STR loci have high degrees of polymorphisms, which may be useful in in- dividual forensic identification and parentage testing in forensic practice.     

云南玉溪汉族15个STR基因座多态性及遗传关系分析

目的调查玉溪汉族人群15个STR基因座的遗传多态性,并分析与国内部分地区汉族群体的遗传关系。方法采用AmpFLSTR Identifiler试剂盒,复合扩增15个STR基因座,计算基因频率及法医学参数;收集国内其他10个群体的遗传学资料进行遗传距离和聚类分析。结果玉溪汉族群体15个STR基因座等位基因及基因型分布符合Hardy-Weinberg平衡定律,PD值在0.790 6～0.968 1之间,PE值在0.315 9～0.733 5之间,PIC值在0.554 6～0.856 4之间,15个基因座累积个体识别力为0.999 999 999 999 999 99,累积非父排除率为0.999 998。不同地区汉族群体间遗传距离分析提示,玉溪汉族与成都汉族遗传距离最近(0.004 0),其次是河南(0.004 5)和潮汕(0.004 7);内蒙古最远(0.036 1)。结论云南玉溪汉族15个STR基因座具有较高的遗传多态性,适于该群体的法医学应用,遗传关系分析结果可为该群体的起源、迁徙及与其他群体的遗传关系分析提供参考。     

新一代遗传标记——InDel研究进展

法医DNA分型经历了三代遗传标记的研究,短串联重复序列(short tandem repeat,STR)作为比较成熟的工具已被广泛运用于法医生物学鉴定中。进一步对人类基因组的探索,先后发现了单核苷酸多态性(single nucleotide polymorphism,SNP)、插入/缺失(Insertion/Deletion,InDel)等一系列遗传标记,而其中InDel作为新型的遗传标记,基本兼具各类遗传标记的优点,受到包括医学分子生物学和法医生物学在内各领域的广泛关注。本文就InDel的研究历史与相应的成果进行简单总结与回顾,以时间轴与研究目的为主要的分类指标,重点关注以多个InDel联合作为遗传标记的多重扩增系统(常染色体或X染色体)在法医生物学及人类学研究中的进展,并对今后该领域研究的方向与暨待解决的问题进行了综述。