用ESI-TOF-MS分型技术检测线粒体DNA高变区碱基组成的多态性

目的 用ESI-TOF-MS分型技术检测线粒体DNA的D环高变区,通过碱基组成分析其多态性.方法 在PLEX-ID技术平台上,分别对mtDNA高变区1(HVⅠ,15924-16428nt)和mtDNA高变区Ⅱ(HVⅡ,31-576 nt)进行碱基组成分析,考察mtDNA在华东汉族人群的多态性,并将该技术应用于一例特殊的亲子鉴定案件.结果 用ESI-TOF-MS分型技术检测线粒体DNA,在高变区Ⅰ的8个区段检见碱基组成的多态性,在mtDNA高变区Ⅱ的10个区段检见多态性.在所应用的亲子鉴定案例中,线粒体DNA标记成了常染色体STR基因座的重要补充,经高变区Ⅰ和高变区Ⅱ的碱基组成检测,最后排除了非母.结论 ESI-TOF-MS检测mtDNA的技术具有良好的应用前景,在一些特殊的案件中,该法可为最终获得可靠鉴定结论提供技术支撑.     

联合应用线粒体SNP和线粒体DNA高变区测序排除同一母系鉴定1例

1 案 例 1.1 简要案情 因申报户口需要,养父母携养女来本所,要求对养父母与养女之间是否存在亲生血缘关系进行鉴定.鉴于养父母各有一个姐姐,还要求对养父母与养女是否来源于同一母系进行鉴定.     

中国汉族人群的线粒体DNA控制区多态性研究

探讨mtDNA多态性在法庭科学中个体识别的理论基础。应用PCR扩增产物直接测序方法 ,对 111名中国北方地区汉族人群无血缘关系个体的mtDNA控制区 (HVⅠ和HVⅡ )进行测序分析。在高变区Ⅰ 15 998～ 16 40 0之间发现 10 2处碱基变异 ,10 3个mtDNA单倍型 ;在高变区Ⅱ 0 0 0 35～ 0 0 36 9之间的发现 36处碱基变异 ,6 9个mtDNA单倍型。其可变碱基的变异形式主要为碱基替代 (转换和颠换 )、插入和缺失 ;碱基转换 (78 9% )明显高于颠换(14 3% )、插入 (3 4% ) ,缺失 (3 4% )。分析表明 ,人群个体mtDNA控制区碱基序列 ,基因多样性为 99 9% ,两个无关个体的偶合概率为 0 92 % ,具有高度序列的多态性     

北京汉族群体17个Y-STR基因座遗传多态性

本文调查了北京地区汉族无关个体DYS456、DYS389I、DYS390、DYS389II、DYS458、DYS19、DYS393、DYS391、DYS439、DYS635、DYS392、Y-GATAH4、DYS437、DYS438、DYS448DYS385a／b等17个Y—STR基因座的遗传多态性,以期为法医学个人识别和亲缘鉴定提供数据。     

线粒体DNA研究进展

人类细胞内存在两套基因组 ,一套是细胞核内的基因组 ,即核ＤＮＡ(ｎｕｃｌｅａｒＤＮＡ ,ｎＤＮＡ) ;另一套是位于细胞质线粒体内的基因组 ,即线粒体ＤＮＡ(ｍｉｔｏｃｈｏｎｄｒｉａｌＤＮＡ ,ｍｔＤＮＡ)。由于线粒体在生命活动中的重要作用及其基因组自身特点 ,使得ｍｔＤＮＡ在细胞遗传学、分子遗传学、发育遗传学和法庭科学等领域受到了广泛重视。一、线粒体ＤＮＡ概述(一 )线粒体ＤＮＡ的结构线粒体ＤＮＡ呈闭环双链结构 ,由 16 5 6 9ｂｐ组成 ,外环为重 (Ｈ)链 ,内环为轻 (Ｌ)链 ,两条链都有编码功能[1] ,共含有 37个基因。ｍｔＤＮ…     

线粒体DNA研究进展

人类细胞内存在两套基因组,一套是细胞核内的基因组,即核DNA(nuclear DNA,nDNA);另一套是位于细胞质线粒体内的基因组,即线粒体DNA(mitochondrial DNA,mtDNA).由于线粒体在生命活动中的重要作用及其基因组自身特点,使得mtDNA在细胞遗传学、分子遗传学、发育遗传学和法庭科学等领域受到了广泛重视.     

北京汉族群体9个STR位点的频率分布及法医学应用

提供北京汉族群体9个STR基因座的频率分布资料，了解其在法医学中的应用价值。应用PCR技术对9个STR基因座分3组进行复合扩增，经PAG电泳分离、银染，扫描仪扫描，计算机判读并保存结果，对北京地区汉族无关个体9个基因座的基因频率分布进行调查。结果显示，上述9个基因座的杂合度为0．6419～0．8092，多态性信息总量为0．9999，鉴别机率为0．9999，匹配机率为2．0×10-9和非父排除率为0．9985。STR3组9个基因座的综合检验可应用于法医学个体识别和亲子鉴定，并达到同一认定的标准。     

线粒体DNA分型技术

综述了线粒体DNA的遗传特性,线粒体DNA分型技术发展历程以及线粒体DNA分型技术相关质量控制、命名原则、异质型影响、结论诠释、数据统计,并展望了线粒体DNA分型技术的未来发展.     

河北汉族人群线粒体DNA HV3(CA)n重复子长度多态性

线粒体DNA的控制区具有高度多态性和母系遗传等特征被广泛应用于法医学个人识别和母系亲权鉴定。目前法医学主要应用HV1、HV2区(16024~16365nt、73~340nt)序列多态性,在m tDNA 00438~00574nt的HV3区域也具有多态性[1],其中位于00514~00524的(CA)n重复子是一个长度多态性区域,重     

武汉地区汉族人群线粒体DNA Region V区9bp片段缺失多态性

目的 研究武汉地区汉族群体线粒体DNA RegionV区9bp片段缺失多态性.方法 PCR扩增后采用银染技术分离片段,检测武汉地区汉族群体线粒体DNA RegionV区9bp片段缺失的频率.结果 在武汉地区汉族239个无关个体中发现标准型、缺失型两种多态类型,9bp片段缺失频率为17.15％.结论 武汉地区汉族群体在DNA RegionV区9bp片段存在缺失多态性.     

Mitochondrial DNA regions HVI and HVII population data.

Data from 1393 unrelated individuals have been compiled from eight population groups: African Americans, Africans (Sierra Leone), U.S. Caucasians, Austrians, French, Hispanics, Japanese, and Asian Americans. The majority of the mtDNA sequences were observed only once within each population group (i.e., ranging from a low of 60.3% (35/58) of the Asian American sequences to a high of 85.3% (93/109) of the French sequences). Genetic diversity ranged from 0.990 in the African sample to 0.998 in African Americans. Random match probability ranged from 2.50% in the Asian American sample to 0.52% in U.S. Caucasians. The average number of nucleotide differences between individuals in a database is greatest for the African American and African samples (14.1 and 13.1, respectively), and the least variable are the Caucasians (ranging from 7.2 to 8.4). Substitutions are the predominate polymorphism, and at least 92% of the substitutions are transitions. The most prevalent transversions are As substituted for Cs and Cs substituted for As. For most population groups these transversions occurred predominately in the HVI region; however, the African, African American, and Hispanic samples also demonstrated a large portion of their C to A and A to C transversions in the HVII region (at sites 186 and/or 189). Most insertions occur in the HVII region at sites 309.1 and 315.1, within a stretch of C's. Insertions of an additional C are common in all population groups. The sequence data were converted to SSO mtDNA types and compared with population data on Caucasians, Africans, Asians, Japanese, and Mexicans described by Stoneking et al. [M. Stoneking, D. Hedgecock, R.G. Higuchi, L. Vigilant, H.A. Erlich, Population variation of human mtDNA control region sequences detected by enzymatic amplification and sequence-specific oligonucleotide probes, Am. J. Hum. Genet. 48 (1991) 370-382] using an R x C contingency table test. Differences between major population groups (i.e., between African, Caucasian, and Asian) are quite evident, and similar ethnic population groups carried similar SSO polymorphism frequencies. There were only a few SSO types that showed significant differences between subpopulation groups. The SSO data alone can not be used to describe the population genetics with complete sequence data. However, the results of the SSO comparisons are similar to other analyses, and differences in sequence data in regions HVI and HVII are greater between major population groups than between subgroups.     

Mitochondrial DNA control region polymorphism in the population of Alagoas state, north-eastern Brazil

The sequences of the two hypervariable (HV) segments of the mitochondrial DNA (mtDNA) control region were determined in 167 randomly selected, unrelated individuals living in the state of Alagoas, north-eastern Brazil. One hundred and forty-five different haplotypes, associated with 139 variable positions, were determined. More than 95% of the mtDNA sequences could be allocated to specific mtDNA haplogroups according to the mutational motifs. Length heteroplasmy in the C-stretch HV1 and HV2 regions was observed in 22 and 11%, respectively, of the population sample. The genetic diversity was estimated to be 0.9975 and the probability of two random individuals presenting identical mtDNA haplotypes was 0.0084. The most frequent haplotype was shared by six individuals. All sequences showed high-quality values and phantom mutations were not detected. The diversity revealed in the mitochondrial control region indicates the importance of this locus for forensic casework and population studies within Alagoas, Brazil.     

Mitochondrial DNA control region population data from Macedonia

Mitochondrial DNA sequences of the entire control region were analyzed in 200 unrelated individuals from Macedonia. A total of 163 different haplotypes were found as determined by 177 polymorphic sites. The probability of a random match was calculated as 1:121 (0.83%). The basic phylogenetic structure of the Macedonian population as derived from its haplogroup distribution is in agreement with other West-Eurasian populations. Upon publication, the population data are going to be available in the EMPOP database (www.empop.org) [W. Parson, A. Dür, EMPOP—a forensic mtDNA database, FSI:Genetics 1 (2) (2007) 88–92; W. Parson, A. Brandstätter, A. Alonso, N. Brandt, B. Brinkmann, A. Carracedo, et al., The EDNAP mitochondrial DNA population database (EMPOP) collaborative exercises: organisation, results and perspectives, Forensic Sci. Int. 139 (2–3) (2004) 215–226.].     

凝血因子B的遗传多态性

目的首次调查北京地区人群体凝血因子B(FB)DNA水平的遗传多态性。方法应用PCR-STR技术。结果推算出FBDNA水平的基因分布频率,符合Hard-Weinberg平衡定律。结论对法医的办案工作,具有一定的实用性和推广意义。     

中国蒙古族群体mtDNA测序的聚类分析及其法医学意义

目的建立一种既节省模板、又能延长测序长度的m tDNA单倍型(群)分析方法,构建中国蒙古族mtDNA单倍型类型关系树。方法用复合扩增、巢式PCR对201名中国蒙古族m tDNA样本进行D-环区、3010~3460、4640~5204、10171~10659和14478~15204编码区域的测序分析,部份样品进行L3953/H4508等区域的测序;根据其多态界定各样本单倍型并进行聚类分析。结果L15996/H107等巢式PCR扩增产物经测序检验结果互不干扰,其分型以A等东亚人群常见的单倍型(群)为主,包括部份HV、K、J、I和U等欧洲人群优势单倍型(群),23个单倍群和共53个单倍型全部归为欧亚人群特有的M和N两大类单倍类群并呈巢式聚类。结论本研究选取的测序区域适用于构建我国各民群的m tDNA单倍型(群);复合扩增、巢式PCR法既节省模板DNA,又延长测序的长度,适用于法医学、考古学研究中的微量样本的检测。     

凝血因子ⅩⅢ B的遗传多态性

目的首次调查北京地区人群体凝血因子ⅩⅢ B(FⅩⅢ B)DNA水平的遗传多态性.方法应用PCR-STR技术.结果推算出FⅩⅢ B DNA水平的基因分布频率,符合Hard-Weinberg平衡定律.结论对法医的办案工作,具有一定的实用性和推广意义.     

Study of the high polymorphism STR locus-FGA in Chinese population

Population genetic study of the complicated STR locus-FGA was performed by the method of PCR in Han population samples. A total of 21 different alleles including 7 interalleles and 63 different genotypes were observed in 349 unrelated individuals. The observed genotype discrimination showed no significant deviation from Hardy-Weinberg equilibrium and 51 family studies showed no mutation. According to the results obtained in this study(DP = 0.9612, PE = 0.7007, PIC = 0.8333), this system can be used as an useful means in forensic identification of both criminal and paternity case.     

Sequence polymorphism of mitochondrial D-loop DNA in the Taiwanese Han population

In order to demonstrate the sequence diversity of mitochondrial D-loop DNA in the Taiwanese Han population, we established a database of 155 unrelated individuals. For each individual, the complete 980bp DNA region from the 5' end of HVI to 3' end of HVII segment was sequenced. In these 155 sequence data, 149 different haplotypes were observed, amongst these haplotypes, 144 were unique, 4 were found in 2 individuals and 1 was found in 3 individuals. When compare to the Anderson sequence, 144 transitions, 24 transversions, 5 insertions and 5 deletions were found. Eight positions exhibited more than one polymorphic sequence, six exhibited two variants while two exhibited three variants. Over the 1024bp that was analysed, pairwise differences between the sequences were 11.35+/-3.53bp. The sequence and nucleotide diversity were 0.9994 and 0.0116, respectively. The probability of two individuals randomly matching over the entire control region was 0.007. The diversity in the mitochondrial D-loop indicates the value of this locus for casework within Taiwan.     

Mitochondrial DNA control region sequence polymorphism in four indigenous tribes of Chotanagpur plateau, India

The analysis of genetic variation, in the nucleotide sequences of mitochondrial DNA, provides unique information in tracing of maternal lineage, determination of population diversity, pharmacogenomics and human identification. This study characterizes the HVR-I and II sequence polymorphism in 80 tribal individuals, belonging to the Austro-Asiatic linguistic family of Chotanagpur plateau, India. A total of 115 polymorphic sites were observed in the sequenced regions and 77 unique haplotypes could be identified.