本文關鍵詞： HLA-B基因 SNP PCR-SSOP PCR-RFLP 親子鑒定　出處：《中國醫(yī)科大學》2005年博士論文　論文類型：學位論文

【摘要】：目的 自二十世紀60年代以來,法醫(yī)學界即開始應用微量淋巴細胞毒試驗和混合淋巴細胞培養(yǎng)法檢測HLA抗原多態(tài)性進行法醫(yī)學親子鑒定。隨著分子生物學技術的迅猛發(fā)展和傳統(tǒng)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基因序列多態(tài)性來進行法醫(yī)學親子鑒定和個人識別,這些方法包括序列特異性引物法、序列特異性寡核苷酸探針法、直接測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。同傳統(tǒng)的血清學、細胞學分型相比,DNA分型準確、迅速、可靠,可避免交叉反應,在法醫(yī)學鑒定中發(fā)揮了重要作用。 HLA-B基因是主要的Ⅰ類基因,序列中SNP位點密集,目前其等位基因數(shù)已經(jīng)達到661種,是多態(tài)性最高的基因,在法醫(yī)學中有較大的應用價值。目前尚缺乏HLA-B特異性SNP位點在中國北方漢族人群中的頻率分布資料以及相關法醫(yī)學應用研究,如果能以特異、靈敏的方法對HLA-B基因序列中的SNP位點進行高通量的分析,獲得其群體分布資料,則必將在混合樣品檢測、非創(chuàng)傷性產(chǎn)前父權鑒定等領域獲得重大應用。 因此,本研究的目的是設計出一組HLA-B基因特異的SNP位點探針,應用PCR-SSOP法和PCR-RFLP法對中國北方漢族人群進行頻率和家系遺傳規(guī)律調(diào)查,獲得相關多態(tài)性數(shù)據(jù);并將探針應用于原位雜交技術中進行非創(chuàng)傷性產(chǎn)前父權鑒定的嘗試,為HLA-B基因多態(tài)性的進一步應用奠定基礎。 方法 本研究根據(jù)已知HLA-B基因序列和各等位基因堿基變異及頻率分布的資料,設計2條HLA-B序列特異性DNA探針B009-Z(序列為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 偽, 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
【學位授予單位】：中國醫(yī)科大學
【學位級別】：博士
【學位授予年份】：2005
【分類號】：D919.1

【引證文獻】

相關期刊論文 前2條

1 王雅莉;曾昭書;李t，

本文編號：1490346