發(fā)布時間：2019-05-23 04:33

【摘要】：研究背景Y-STR分型在法醫(yī)學(xué)檢案中已得到廣泛的應(yīng)用,尤其在性犯罪案件調(diào)查、父系親緣關(guān)系鑒定及現(xiàn)場生物檢材的群體來源推斷等方面發(fā)揮了獨(dú)特的作用。但由于父系遺傳的特點(diǎn),Y-STR在部分地區(qū)和人群多態(tài)性較差,傳統(tǒng)的Y-STR分型試劑盒不具備足夠的個人分辨力,致使其應(yīng)用價值受到限制。此外,目前使用的Y-STR基因座缺乏區(qū)分來自同一父系男性個體的能力,除非發(fā)生突變,同一父系男性個體具有同樣的Y-STR分型,不能從個體層面進(jìn)行有效區(qū)分。近年來,RM Y-STR的提出有望解決Y-STR分辨力有限和同一父系男性個體區(qū)分的難題。目的調(diào)查13個RM Y-STR基因座在湖北漢族人群的等位基因分布和多態(tài)性,構(gòu)建包含13個基因座的熒光復(fù)合擴(kuò)增體系,并針對父-子樣本調(diào)查漢族人群的突變率。方法1.收集252個無關(guān)男性個體血液樣本分別進(jìn)行13個RM Y-STR基因座檢測,結(jié)合片段測序明確各基因座的重復(fù)結(jié)構(gòu)并進(jìn)行等位基因命名,分析各基因座的基因多態(tài)性和單倍型頻率;2.根據(jù)等位基因片段長度范圍和復(fù)合擴(kuò)增引物設(shè)計(jì)要求,重新設(shè)計(jì)引物并標(biāo)記四色熒光染料,構(gòu)建13個RM Y-STR基因座熒光復(fù)合擴(kuò)增體系;3.收集父-子樣本,使用該復(fù)合擴(kuò)增體系進(jìn)行突變率調(diào)查,評估其在漢族人群中的法醫(yī)學(xué)應(yīng)用價值。結(jié)果1.13個RM Y-STR基因座的重復(fù)結(jié)構(gòu)和等位基因命名得到確認(rèn),各基因座在漢族人群中多態(tài)性均良好,GD或HD變化范圍為0.7862~0.9953;2.本研究構(gòu)建的13個RM Y-STR基因座熒光復(fù)合擴(kuò)增體系擴(kuò)增效果良好,分型穩(wěn)定,可重復(fù)性好;3.13個RM Y-STR基因座在湖北漢族人群中父-子分辨率為19.0%,9個基因座的突變率在10-2以上,與報道的突變率相近;基因座DYF403S1b、DYS526b、DYS570和DYS626突變率較低,為5~7×10-3。結(jié)論13個RM Y-STR基因座在湖北漢族人群中均具備良好的基因/單倍型多樣性,且9個基因座突變率在10-2以上,說明快速突變基因座在漢族人群中的應(yīng)用具有極大的潛力。通過豐富其他地區(qū)的群體數(shù)據(jù),有針對性地調(diào)整基因座組合,可有效發(fā)揮快速突變基因座的應(yīng)用價值。本研究構(gòu)建的13個RM Y-STR基因座熒光復(fù)合擴(kuò)增體系具備分型準(zhǔn)確、穩(wěn)定性強(qiáng)、靈敏度高等特點(diǎn),為快速突變基因座體系在法醫(yī)學(xué)中的實(shí)際應(yīng)用提供了有效參考。
[Abstract]:Background Y-STR typing has been widely used in forensic cases, especially in the investigation of sexual crimes, the identification of patrilineal kinship and the inference of population sources of biological samples on the spot. However, due to the characteristics of patrilineal heredity, the polymorphism of Y-STR in some areas and populations is poor, and the traditional Y-STR typing kit does not have enough personal resolution, which limits its application value. In addition, the Y-STR loci currently used lack the ability to distinguish male individuals from the same patriline. unless there is a mutation, the same patrilineal male individuals have the same Y-STR typing and can not effectively distinguish them from the individual level. In recent years, the proposal of RM Y-STR is expected to solve the problems of limited Y-STR resolution and individual differentiation of the same patrilineal male. Objective to investigate the distribution and polymorphism of 13 RM Y-STR loci in Hubei Han population, to construct a fluorescence compound amplification system containing 13 loci, and to investigate the mutation rate of Han population according to father-son samples. Method 1. The blood samples of 252 unrelated male individuals were collected and 13 RM Y-STR loci were detected. The repeat structures of each locus were identified by fragment sequencing and the alleles were named. The gene polymorphism and haplotype frequency of each locus were analyzed. 2. According to the length range of allelic fragments and the design requirements of compound amplification primers, 13 RM Y-STR loci fluorescence compound amplification systems were constructed by redesigning primers and labeling four-color fluorescent dyes. The father-son samples were collected and the mutation rate was investigated by using the compound amplification system to evaluate the value of forensic medicine in Han population. Results 1. The repeat structure and allelic nomenclature of 13 RM Y-STR loci were confirmed. The polymorphism of each locus was good in Han population, and the range of GD or HD was 0.7862 鈮，

本文編號：2483603