HLA-B基因多态性在法医学中的应用研究

发布时间：2018-02-04 13:41

本文关键词： HLA-B基因 SNP PCR-SSOP PCR-RFLP 亲子鉴定　出处：《中国医科大学》2005年博士论文　论文类型：学位论文

【摘要】：目的自二十世纪60年代以来,法医学界即开始应用微量淋巴细胞毒试验和混合淋巴细胞培养法检测HLA抗原多态性进行法医学亲子鉴定。随着分子生物学技术的迅猛发展和传统抗原分型方法的局限,目前已主要采用基因分型方法分析HLA基因序列多态性来进行法医学亲子鉴定和个人识别,这些方法包括序列特异性引物法、序列特异性寡核苷酸探针法、直接测序分型法以及基因芯片法等,所涉及的基因包括HLA-A、HLA-DRB1以及HLA-DQα等。同传统的血清学、细胞学分型相比,DNA分型准确、迅速、可靠,可避免交叉反应,在法医学鉴定中发挥了重要作用。 HLA-B基因是主要的Ⅰ类基因,序列中SNP位点密集,目前其等位基因数已经达到661种,是多态性最高的基因,在法医学中有较大的应用价值。目前尚缺乏HLA-B特异性SNP位点在中国北方汉族人群中的频率分布资料以及相关法医学应用研究,如果能以特异、灵敏的方法对HLA-B基因序列中的SNP位点进行高通量的分析,获得其群体分布资料,则必将在混合样品检测、非创伤性产前父权鉴定等领域获得重大应用。因此,本研究的目的是设计出一组HLA-B基因特异的SNP位点探针,应用PCR-SSOP法和PCR-RFLP法对中国北方汉族人群进行频率和家系遗传规律调查,获得相关多态性数据;并将探针应用于原位杂交技术中进行非创伤性产前父权鉴定的尝试,为HLA-B基因多态性的进一步应用奠定基础。方法本研究根据已知HLA-B基因序列和各等位基因碱基变异及频率分布的资料,设计2条HLA-B序列特异性DNA探针B009-Z(序列为5’-
[Abstract]:Purpose Since 1960s. The medical profession began to use microlymphocytotoxicity test and mixed lymphocyte culture method to detect the polymorphism of HLA antigen in forensic medicine. With the rapid development of molecular biology technology and traditional antigen typing prescription. The limitations of law. At present, genetic typing method has been mainly used to analyze the polymorphism of HLA gene for paternity identification and personal identification in forensic medicine. These methods include sequence specific primer method and sequence specific oligonucleotide probe method. Direct sequencing typing method and gene chip method, the genes involved, including HLA-An HLA-DRB1 and HLA-DQ 伪, compared with the traditional serological and cytological typing. DNA typing is accurate, rapid, reliable, can avoid cross-reaction, and plays an important role in forensic identification. HLA-B gene is the main class I gene. The SNP loci are dense in the sequence. At present, the number of alleles has reached 661.It is the most polymorphic gene. At present, there is a lack of frequency distribution data of HLA-B specific SNP locus in Han population in northern China, and the relevant forensic application research, if it can be specific. The high throughput analysis of SNP loci in HLA-B gene sequence by a sensitive method, and the data of population distribution, will certainly be detected in mixed samples. Non-traumatic prenatal paternity identification and other fields have been important applications. Therefore, the aim of this study is to design a set of HLA-B gene specific SNP locus probes. PCR-SSOP and PCR-RFLP methods were used to investigate the frequency and family genetic rules of Han population in northern China, and relevant polymorphism data were obtained. The probe was applied to non-traumatic prenatal paternity identification in situ hybridization, which laid a foundation for the further application of HLA-B gene polymorphism. Method This study was based on the known HLA-B gene sequence, allele base variation and frequency distribution. Design of two HLA-B sequence-specific DNA probes B009-Z5
【学位授予单位】：中国医科大学
【学位级别】：博士
【学位授予年份】：2005
【分类号】：D919.1

【引证文献】