贵州汉族人群22个STR多态性的调查及11个SNPs复合检测体系的构建

发布时间：2018-05-01 07:11

本文选题：单核苷酸多态性 + 个体识别　；参考：《贵州医科大学》2017年硕士论文

【摘要】：目的:本研究旨在调查贵州地区汉族人群22个STR的遗传多态性、观察分析突变特点,对其系统效能进行评估。人类基因组数据库中,按照个体识别单核苷酸多态性(Individual Identification single nucleotide polymorphisms,IISNPs)的标准筛选出满足要求的单核苷酸多态性(single nucleotide polymorphisms,SNPs)位点,通过单碱基延伸(Single base extension,SBE)反应与毛细管电泳检测技术构建SNPs位点复合检测体系,为现有STR和SNP检验体系作补充。方法:1.应用PowerPlex?Fusion试剂对2018份贵州地区汉族无关个体进行22个常染色体STR的等位基因分型,统计贵州汉族人群等位基因频率、法医学参数、突变位点及频率。2.在SNP数据库里选择20个符合要求的SNP位点,根据公布的基因序列,应用引物设计软件Primer 5.0自行设计PCR引物,使用DNAMAN软件自行设计单碱基延伸反应引物。3.构建复合PCR、复合单碱基延伸反应检测体系,应用ABI 3500遗传分析仪对SNPs位点进行等位基因分型,3500 Data Collection V3.0软件搜集数据,GeneMapper ID-X 1.4软件分析结果。4.应用构建的SNPs复合检测体系调查103份贵州地区汉族无关个体的遗传多态性,并对该SNPs复合检测体系在亲权鉴定中的应用价值进行评估。5.应用构建的SNPs复合检测体系检测5例贵州地区具有亲缘关系的汉族二代家系鉴定样本,分析其是否符合孟德尔遗传规律。6.应用该体系对一例22个STR分型突变累积父权指数不达104的案例进行11个SNPs遗传标记的补充,统计其累积父权指数。结果:1.获得贵州地区2018份汉族无关个体22个常染色体STR位点的群体遗传学数据,该抽样调查的人群基因型频率分布符合Hardy-Weinberg平衡。22个STR位点累积个人识别能力(CPD)为1-2×10-26,累积非父排除概率(CPE)为0.999 999 999,累积匹配概率(CMP)为1.1516×10-26。2.自行设计常染色体SNPs位点的PCR引物及单碱基延伸反应引物,在20个SNPs位点中筛选出11个SNPs位点构建复合检测体系,获得103份贵州地区汉族无关个体11个SNPs位点的等位基因频率及法医学参数。3.5例贵州地区汉族二代家系鉴定样本,均符合孟德尔遗传规律。4.应用该体系对一例STR分型突变累积父权指数不达104的亲子鉴定案例分析,肯定其亲权关系。结论:1.本文研究的D1S1656等22个STR基因座,在贵州地区汉族人群中具备良好的遗传多态性,可在法医亲子鉴定和个体识别中应用。2.成功构建含11个IISNPs位点的复合检测体系,可作为现有STR检测体系和SNP位点的补充。
[Abstract]:Objective: to investigate the genetic polymorphisms of 22 STR in Guizhou Han population and analyze the mutation characteristics and evaluate its systematic effectiveness. In the human genome database, single Identification single nucleotide polymorphisms (SNPs) were screened according to individual identification criteria for individual Identification single nucleotide polymorphisms (SNPs). The compound detection system of SNPs locus was constructed by single base extension (SBE) reaction and capillary electrophoresis (CE) to supplement the existing STR and SNP detection systems. Method 1: 1. The alleles of 22 autosomal STR alleles of 2018 unrelated individuals of Han nationality in Guizhou were classified with PowerPlex?Fusion reagent. The allelic frequencies, forensic parameters, mutation sites and frequencies of 22 autosomal STR alleles in Guizhou Han population were analyzed. Twenty SNP loci were selected from SNP database. According to the published gene sequence, PCR primers were designed by Primer 5.0 and single base extension primers .3were designed by DNAMAN software. ABI 3500 genetic analyzer was used to type the alleles of SNPs loci. The data were collected by using the software of 3 500 Data Collection V3.0. The results of GeneMapper ID-X 1.4 software were analyzed by using GeneMapper ID-X 1.4 software. The genetic polymorphisms of 103 unrelated individuals of Han nationality in Guizhou were investigated by using the constructed SNPs complex detection system, and the application value of the SNPs complex detection system in paternity test was evaluated. The constructed SNPs complex detection system was used to detect the identification samples of the second generation Han families in Guizhou area, and to analyze whether it was in accordance with the Mendelian heredity law. 6. This system was used to supplement 11 SNPs genetic markers in a case with 22 STR genotyping mutations whose cumulative paternity index was not up to 104, and its cumulative paternity index was counted. The result is 1: 1. Population genetic data of 22 autosomal STR loci in 2018 unrelated individuals of Han nationality in Guizhou were obtained. The genotype frequency distribution in this sample was consistent with the Hardy-Weinberg balance. The cumulative personal recognition ability of 22 STR loci was 1-2 脳 10-26, the cumulative non-paternal exclusion probability was 0.999 9999, and the cumulative matching probability was 1.1516 脳 10-26.2. PCR primers and single base extension primers for autosomal SNPs loci were designed and 11 SNPs loci were screened out of 20 SNPs loci to construct a complex detection system. The allelic frequencies of 11 SNPs loci and forensic medical parameters were obtained from 103 unrelated individuals of Han nationality in Guizhou. The identification samples of second-generation families of Han nationality in Guizhou were all in accordance with the Mendelian genetic rule .4. The system was used to analyze a case of paternity test in which the cumulative paternity index of STR mutation was not up to 104, and the paternity relationship was confirmed. Conclusion 1. The 22 STR loci studied in this paper have good genetic polymorphisms in Guizhou Han population and can be used in forensic paternity test and individual identification. A complex detection system containing 11 IISNPs loci was successfully constructed and can be used as a complement to existing STR detection systems and SNP loci.
【学位授予单位】：贵州医科大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：D919

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