四個(gè)X-STR基因座熒光復(fù)合擴(kuò)增體系的建立及其法醫(yī)遺傳學(xué)研究
發(fā)布時(shí)間：2018-08-17 13:53
【摘要】： 目的:本課題旨在建立四個(gè)x染色體特異性短串聯(lián)重復(fù)基因座DXS9902、DXS6800、DXS6799和DXS7132的熒光復(fù)合擴(kuò)增體系并對(duì)其進(jìn)行法醫(yī)學(xué)實(shí)用性研究;利用所建立的體系調(diào)查北京漢族人群中這四個(gè)基因座的遺傳多念性,為DNA數(shù)據(jù)庫(kù)建立和群體遺傳學(xué)提供基礎(chǔ)數(shù)據(jù)。 方法:對(duì)國(guó)內(nèi)外文獻(xiàn)中查到的8個(gè)已有報(bào)道的X-STR基因座進(jìn)行篩選,從中獲得四個(gè)多態(tài)性好、個(gè)人識(shí)別率高,擴(kuò)增產(chǎn)物長(zhǎng)度片段有區(qū)別的基因座,利用傳統(tǒng)的復(fù)合擴(kuò)增法,ABI PRISM 3100毛細(xì)管電泳進(jìn)行熒光標(biāo)記復(fù)合擴(kuò)增的研究,并對(duì)復(fù)合擴(kuò)增體系的靈敏度和準(zhǔn)確度,不同擴(kuò)增參數(shù)下復(fù)合擴(kuò)增的效果進(jìn)行研究及法庭科學(xué)有效性驗(yàn)證,并應(yīng)用于實(shí)際案件;用所建立的體系對(duì)北京漢族200名無(wú)關(guān)男性個(gè)體及200名無(wú)關(guān)女性個(gè)體進(jìn)行群體遺傳學(xué)調(diào)查。 結(jié)果:建立了DXS9902、DXS6800、DXS6799和DXS7132的熒光標(biāo)記復(fù)合擴(kuò)增體系,該體系條件穩(wěn)定,結(jié)果準(zhǔn)確可靠。法庭科學(xué)有效性研究發(fā)現(xiàn),應(yīng)用該體系可獲得準(zhǔn)確分型結(jié)果的最小模板量為0.25 ng,對(duì)豬,牛,羊,雞,魚(yú)和鴨等常見(jiàn)動(dòng)物檢測(cè)均未得到特異性分型結(jié)果,同一個(gè)體血,骨骼肌,心肌,肝臟,肺臟和腦等組織分型結(jié)果一致。對(duì)實(shí)際案件中血痕、軟骨,煙蒂,精斑,毛發(fā)等常見(jiàn)檢材進(jìn)行檢測(cè),均得到可靠分型結(jié)果。北京漢族人群調(diào)查得到四個(gè)基因座的基因頻率(0.002—0.817);個(gè)人識(shí)別率(0.526-0.886);非父排除率(0.32-0.691);四個(gè)基因座獨(dú)立性分析,未檢測(cè)到連鎖不平衡。 結(jié)論:DXS9902、DXS6800、DXS6799和DXS7132的熒光標(biāo)記復(fù)合擴(kuò)增體系,分型結(jié)果穩(wěn)定可靠,具有高度準(zhǔn)確性,高靈敏度,種屬特異性、組織同一性、檢材適應(yīng)性;該方法簡(jiǎn)便、高效,易于掌握,便于推廣。群體遺傳學(xué)調(diào)查表明,該系統(tǒng)個(gè)體識(shí)別能力較強(qiáng),非父排除率較高,基因座之間具有相對(duì)獨(dú)立性,適用于法醫(yī)學(xué)個(gè)人識(shí)別和親子鑒定。
[Abstract]:Objective: to establish the fluorescent multiplex amplification system of DXS9902 and DXS6800 DXS6799 and DXS7132 for four X-chromosome specific short tandem repeat loci (DXS9902DXS6799 and DXS7132) and to study the forensic practicability of DXS9902DXS6799 and DXS6799. The genetic polymorphisms of these four loci in the Han population of Beijing were investigated by using the established system to provide basic data for the establishment of DNA database and population genetics. Methods: eight reported X-STR loci were screened in the literature at home and abroad. Four loci with good polymorphism, high personal recognition rate and different amplified product length fragments were obtained. The fluorescence labeling multiplex amplification of ABI PRISM 3100 was studied by using traditional multiplex amplification method, and the sensitivity and accuracy of the system were also studied. The effects of multiplex amplification under different amplification parameters were studied and the validity of forensic science was verified and applied to practical cases. The established system was used to investigate the population genetics of 200 unrelated male individuals and 200 unrelated female individuals of Han nationality in Beijing. Results: the fluorescence labeling multiplex amplification system of DXS9902 DXS6800 and DXS6799 and DXS7132 was established. The conditions of the system were stable and the results were accurate and reliable. Forensic scientific and effective studies showed that the minimum amount of template used to obtain accurate typing results was 0.25 ng. No specific typing results were obtained for common animals, such as pigs, cattle, sheep, chickens, fish and ducks, and the same body blood, skeletal muscle, and so on. The results of myocardial, liver, lung and brain tissue typing were consistent. The blood marks, cartilage, cigarette butts, sperm spots, hair and other common samples were detected in the actual cases, and the reliable typing results were obtained. The gene frequency of four loci (0.002-0.817), personal recognition rate (0.526-0.886), non-paternal exclusion rate (0.32-0.691) and linkage disequilibrium were not detected. Conclusion the fluorescent multiplex amplification system of DXS9902, DXS6800, DXS6799 and DXS7132 is stable and reliable, with high accuracy, high sensitivity, species-specificity, tissue identity and material adaptability. The method is simple, efficient, easy to master and easy to be popularized. Population genetic investigation showed that the system had strong individual recognition ability, high non-paternal exclusion rate and relative independence between loci, and was suitable for forensic personal identification and paternity testing.
【學(xué)位授予單位】：山西醫(yī)科大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2009
【分類(lèi)號(hào)】：D919
本文編號(hào)：2187846
[Abstract]:Objective: to establish the fluorescent multiplex amplification system of DXS9902 and DXS6800 DXS6799 and DXS7132 for four X-chromosome specific short tandem repeat loci (DXS9902DXS6799 and DXS7132) and to study the forensic practicability of DXS9902DXS6799 and DXS6799. The genetic polymorphisms of these four loci in the Han population of Beijing were investigated by using the established system to provide basic data for the establishment of DNA database and population genetics. Methods: eight reported X-STR loci were screened in the literature at home and abroad. Four loci with good polymorphism, high personal recognition rate and different amplified product length fragments were obtained. The fluorescence labeling multiplex amplification of ABI PRISM 3100 was studied by using traditional multiplex amplification method, and the sensitivity and accuracy of the system were also studied. The effects of multiplex amplification under different amplification parameters were studied and the validity of forensic science was verified and applied to practical cases. The established system was used to investigate the population genetics of 200 unrelated male individuals and 200 unrelated female individuals of Han nationality in Beijing. Results: the fluorescence labeling multiplex amplification system of DXS9902 DXS6800 and DXS6799 and DXS7132 was established. The conditions of the system were stable and the results were accurate and reliable. Forensic scientific and effective studies showed that the minimum amount of template used to obtain accurate typing results was 0.25 ng. No specific typing results were obtained for common animals, such as pigs, cattle, sheep, chickens, fish and ducks, and the same body blood, skeletal muscle, and so on. The results of myocardial, liver, lung and brain tissue typing were consistent. The blood marks, cartilage, cigarette butts, sperm spots, hair and other common samples were detected in the actual cases, and the reliable typing results were obtained. The gene frequency of four loci (0.002-0.817), personal recognition rate (0.526-0.886), non-paternal exclusion rate (0.32-0.691) and linkage disequilibrium were not detected. Conclusion the fluorescent multiplex amplification system of DXS9902, DXS6800, DXS6799 and DXS7132 is stable and reliable, with high accuracy, high sensitivity, species-specificity, tissue identity and material adaptability. The method is simple, efficient, easy to master and easy to be popularized. Population genetic investigation showed that the system had strong individual recognition ability, high non-paternal exclusion rate and relative independence between loci, and was suitable for forensic personal identification and paternity testing.
【學(xué)位授予單位】：山西醫(yī)科大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2009
【分類(lèi)號(hào)】：D919
【引證文獻(xiàn)】
相關(guān)碩士學(xué)位論文 前1條
1 王磊;GATA165B12，DXS6800，GATA31E08基因座多態(tài)性及其熒光標(biāo)記復(fù)合擴(kuò)增體系的法醫(yī)學(xué)應(yīng)用研究[D];鄭州大學(xué);2010年
，本文編號(hào)：2187846
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