山西汉族人群DXS7423、DXS6799基因座遗传多态性及法医学应用可行性探讨

发布时间：2018-11-13 11:37

【摘要】： 目的调查山西汉族人群中DXS7423、DXS6799基因座的等位基因频率分布、单倍型频率分布及相关遗传学参数等,为人类X染色体STR基因座群体遗传学数据库的建立提供基础数据,并对其法医学应用的可行性进行探讨。方法(1)样本采集:随机抽取250名山西汉族无关个体(男性154,女性96)静脉血进行两个基因座的多态性分析;采集30例二联体家系血进行突变观察;采集同一女性尸体的血液、肌肉、心脏、肝脏、肾脏、皮肤组织进行同一性观察。(2)方法:基因组DNA用酚-氯仿有机溶剂法提取,经PCR扩增后,用8%非变性聚丙烯酰胺凝胶电泳,硝酸银染色,将各等位基因电泳谱带与等位基因Ladder进行比对分;命名根据国际法医血液学会(ISFH)DNA委员会推荐的命名原则进行。(3)数据处理:用直接计算法计算等位基因频率、基因型频率和单倍型频率;用χ2检验进行男女群体等位基因频率差异比较、女性样本Hardy-Weinberg平衡吻合度检验和不同群体等位基因频率差异比较,并计算各种相关遗传学参数。结果(1)山西汉族250名无关个体中,DXS7423基因座检出4个等位基因13～16,基因频率分布范围0.0057～0.5029;DXS6799基因座检出7个等位基因9～15,基因频率分布范围0.0058～0.4017。χ2检验结果表明男女性人群之间两个基因座的等位基因频率分布均无显著性差异(P0.05),因此可以合并男女数据进行总体等位基因频率的统计。 DXS7423、DXS6799基因座在山西汉族96名无关女性个体中分别检出7、13种基因型,两基因座女性基因型频率分布均符合Hardy-Weinberg平衡(P0.05)。两个基因座女性个人识别率分别为0.8020、0.8963;非父排除率分别为0.3425、0.4973;男性个人识别率分别为0.5767、0.7428;合并数据后得各基因座的多态性信息含量分别为0.6795、0.7643。在154名无关男性个体中两个基因座组成的单倍型观察到19种,单倍型多样性、个人识别率、非父排除率分别为0.8890、0.8890、0.7947。 (2)个体组织同一性:同一女性尸体不同组织DNA分型结果表明,两基因座在同一个体中均具有组织同一性,符合STR基因座法医学应用标准。 (3)遗传稳定性: 30例已经确定父女关系的两代家系中,父亲的X染色体均能稳定地遗传给女儿,符合孟德尔遗传规律。结论DXS7423、DXS6799基因座在山西汉族人群中均具有较高的遗传多态性和遗传稳定性,在法医学应用、人类遗传学研究等领域中具有重要价值。
[Abstract]:Objective to investigate the allele frequency distribution, haplotype frequency distribution and related genetic parameters of DXS7423,DXS6799 locus in Shanxi Han population, so as to provide basic data for the establishment of population genetic database of human X chromosome STR locus. The feasibility of its forensic application was discussed. Methods (1) sampling: 250 unrelated individuals of Shanxi Han nationality (male 154, female 96) were randomly selected for polymorphism analysis of two loci, and 30 cases of disjoint blood were collected for mutation observation. Blood, muscle, heart, liver, kidney and skin tissues of the same female cadaver were collected for identity observation. (2) Genomic DNA was extracted by phenol-chloroform organic solvent method and amplified by PCR. The allele electrophoresis bands were compared with allele Ladder by 8% non-denatured polyacrylamide gel electrophoresis and silver nitrate staining. The nomenclature was carried out according to the naming principle recommended by the (ISFH) DNA Committee of the International Society of Forensic Medicine. (3) data processing: the allele frequency, genotype frequency and haplotype frequency were calculated by direct calculation; The allele frequency difference of male and female population was compared by 蠂 2 test. The Hardy-Weinberg equilibrium coincidence test and the allele frequency difference of different populations were compared in female samples. The genetic parameters were calculated. Results (1) in 250 unrelated individuals of Shanxi Han nationality, 4 alleles were detected in the DXS7423 locus, and the frequency of the alleles was 0.0057 / 0.5029; Seven alleles were detected in DXS6799 loci, and the frequency distribution range was 0.0058, 0.4017.The results of 蠂 2 test showed that there was no significant difference in allele frequency distribution between male and female (P0.05). Therefore, the total allele frequency statistics can be combined with male and female data. DXS7423,DXS6799 loci were detected in 96 unrelated female individuals of Shanxi Han nationality, and 13 genotypes were detected respectively. The frequencies of the two loci were in accordance with the Hardy-Weinberg balance (P0.05). The personal recognition rate of the two loci was 0.8020 / 0.8963, the non-paternal exclusion rate was 0.3425 / 0.4973, and the male's personal recognition rate was 0.5767 / 0.7428, respectively. The polymorphic information content of each locus was 0.6795 and 0.7643, respectively. 19 haplotypes composed of two loci were observed in 154 unrelated male individuals. The haplotypes diversity, individual recognition rate and non-paternal exclusion rate were 0.8890 / 0.8890 / 0.7947, respectively. (2) individual tissue identity: the results of DNA typing in different tissues of the same female cadaver showed that the two loci had tissue identity in the same body, which was in accordance with the standard of forensic application of STR locus. (3) genetic stability: in 30 families with established paternal and daughter relationships, the X chromosome of the father could be inherited to his daughter stably, which was in accordance with the Mendelian inheritance law. Conclusion DXS7423,DXS6799 loci have high genetic polymorphism and genetic stability in Shanxi Han population and are of great value in forensic application and human genetics research.
【学位授予单位】：山西医科大学
【学位级别】：硕士
【学位授予年份】：2009
【分类号】：D919

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