本文選題：法醫(yī)遺傳學(xué) + 微小片段短串聯(lián)重復(fù)序列(miniSTR)　； 參考：《四川大學(xué)》2007年碩士論文

【摘要】： 目的miniSTR技術(shù)是當今法醫(yī)物證學(xué)研究高度降解檢材的熱點，本課題旨在研究3個miniSTR基因座(D10S1248，D14S1434和D22S1045)在中國成都漢族人群中的特征，構(gòu)建復(fù)合擴增熒光標記檢測系統(tǒng)及評估其法醫(yī)學(xué)應(yīng)用價值。方法使用Coble的引物，在中國成都200個無關(guān)漢族個體中進行3個miniSTR基因座PCR擴增，擴增產(chǎn)物使用聚丙烯酰胺凝膠電泳分離，銀染法顯色，以獲得群體基本數(shù)據(jù)。建立這3個miniSTR基因座的復(fù)合擴增體系，利用熒光復(fù)合擴增技術(shù)擴增，ABI PRISM~(?) 310基因分析儀對產(chǎn)物進行檢測，GeneScan3.7及Geno Typer3.7軟件分析結(jié)果；并按照美國DNA分析方法技術(shù)工作組(Scientific Working Group on DNA Analysis Methods，，SWGDAM)的指導(dǎo)方案進行法醫(yī)學(xué)應(yīng)用性研究，用不同的組織、不同的介質(zhì)、常見的動物對復(fù)合擴增系統(tǒng)進行測試，用實際案例進行驗證。結(jié)果獲得了3個miniSTR基因座的基因頻率、個人識別率、非父排除率等群體遺傳學(xué)數(shù)據(jù)，在D10S1248基因座觀察到了新的等位基因8，在D14S1434基因座發(fā)現(xiàn)了不同的重復(fù)序列；通過采用復(fù)合擴增方法，建立了D10S1248、D14S1434和D22S1045熒光標記復(fù)合擴增體系。法醫(yī)應(yīng)用性研究證明該體系與12種常見動物和微生物無交叉反應(yīng)；可對2ng的微量DNA模板分型；對同一個體的不同組織檢測結(jié)果一致；能夠?qū)ΤＲ娊橘|(zhì)上的斑痕檢材正確分型；對放置20年的陳舊血痕仍然可分型；應(yīng)用于案例檢測，結(jié)果與鑒定結(jié)論一致。結(jié)論3個miniSTR遺傳標記(D10S1248，D14S1434和D22S1045)在中國人群中有較高的多態(tài)性及較高的個人識別率，在不同的人群中存在遺傳差異。建立了這3個miniSTR基因座的熒光標記復(fù)合擴增體系miniplex，該體系可運用于熒光標記毛細管電泳激光自動分析檢測平臺，具備法醫(yī)學(xué)實踐應(yīng)用的可行性，為分析DNA降解檢材提供了一個有效的途徑。
[Abstract]:Objective to study the characteristics of three miniSTR loci (D10S1248, D14S1434 and D22S1045) in Chengdu, China.To construct a multiplex amplification fluorescent marker detection system and evaluate its forensic application value.Methods the primers of Coble were used to amplify three miniSTR loci from 200 unrelated Han Chinese individuals in Chengdu, China. The amplified products were separated by polyacrylamide gel electrophoresis and colored by silver staining to obtain the basic population data.To establish the multiplex amplification system of the three miniSTR loci, and to detect the products by using the fluorescence multiplex amplification technique to detect the products with GeneScan3.7 and Geno Typer3.7 software.In accordance with the guidance of Scientific Working Group on DNA Analysis methods (SWGDAM), the United States DNA Analytical Methodology Technical working Group conducted forensic application research, and tested the multiplex amplification system with different tissues, different media and common animals.An actual case is used to verify the problem.Results the population genetic data of three miniSTR loci, such as gene frequency, personal identification rate and non-paternal exclusion rate, were obtained. New alleles 8 were observed in D10S1248 loci and different repeat sequences were found in D14S1434 loci.The multiplex amplification system of D10S1248, D14S1434 and D22S1045 was established by using the method of multiplex amplification.It can still be classified for 20 years old blood marks, and the results are consistent with the identification results.Conclusion three miniSTR markers D10S1248, D14S1434 and D22S1045) have higher polymorphism and higher personal identification rate in Chinese population, and there are genetic differences among different populations.The fluorescent multiplex amplification system of these three miniSTR loci was established. The system can be used in fluorescence labeling capillary electrophoresis laser automatic analysis and detection platform, which has the feasibility of forensic practice.It provides an effective way for the analysis of DNA degradation samples.
【學(xué)位授予單位】：四川大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2007
【分類號】：D919

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