GATA165B12,DXS6800,GATA31E08基因座多態(tài)性及其熒光標(biāo)記復(fù)合擴增體系的法醫(yī)學(xué)應(yīng)用研究
本文選題:短串聯(lián)重復(fù)序列 切入點:X單倍型 出處:《鄭州大學(xué)》2010年碩士論文 論文類型:學(xué)位論文
【摘要】: 目的本課題旨在提高X染色體的鑒別能力,為法醫(yī)X染色體分型提供新的遺傳標(biāo)記。依托毛細管電泳激光熒光檢測平臺,運用熒光標(biāo)記復(fù)合擴增技術(shù),對3個X-STR基因座進行片段分析,建立一套新GATA165B12、DXS6800、GATA31E08三個X-STR的基因座復(fù)合擴增體系,為等位基因的命名奠定基礎(chǔ),為法醫(yī)學(xué)應(yīng)用提供概率計算依據(jù)。 方法PCR擴增154名無關(guān)河南漢族女性個體和65名無關(guān)河南漢族男性個體血樣DNA樣本,PCR產(chǎn)物先使用聚丙稀酰胺凝膠電泳分離,EB染色紫外顯像,后用自行設(shè)計的熒光標(biāo)記引物復(fù)合擴增,毛細管電泳熒光自動檢測系統(tǒng)對復(fù)合擴增產(chǎn)物進行檢測和分型,用實際案例進行驗證。 結(jié)果154名河南漢族女性無關(guān)個體和65名河南漢族男性無關(guān)個體在GATA165B12、DXS6800、GATA31E08三個X-STR的基因座分別檢出4、7、12個等位基因,雜合度觀察值分別為0.617、0.266和0.766。154名河南漢族女性無關(guān)個體DP值分別為0.806、0.46和0.906,PIC值分別為0.57、0.28和0.72,PE值分別為0.312、0.051和0.538,三個基因座累計個人識別能力為0.99015,三個基因座累計非父排除概率為0.6984;經(jīng)卡方檢驗,三個STR位點的Hardy-Weinberg平衡概率均大于0.05。65名河南漢族男性無關(guān)個體中檢測28種單倍型,其單倍型GD為0.9529。 結(jié)論對三個X-STR基因座的種屬特異性、組織特異性、靈敏度及實際案件中的應(yīng)用等問題進行研究,結(jié)果表明三個X-STR基因座具有較好的種屬特異性、組織特異性、靈敏度高,應(yīng)用熒光標(biāo)記X-STR復(fù)合擴增檢驗技術(shù)在實際檢案中具有良好效果。
[Abstract]:Objective to improve the ability of X chromosome identification and to provide a new genetic marker for forensic X chromosome typing. By fragment analysis of three X-STR loci, a new amplification system of three X-STR loci of GATA165B12 DXS6800GTA31E08 was established, which laid the foundation for allele naming and provided the basis of probability calculation for forensic application. Methods the PCR products of blood samples from 154 unrelated Henan Han women and 65 unrelated Henan Han Chinese individuals were amplified by polymerase chain reaction (PCR) using polyacrylamide gel electrophoresis to separate EB staining ultraviolet imaging. The multiplex amplification was carried out with self-designed fluorescent labeled primers, and the multiplex amplification products were detected and typed by capillary electrophoresis fluorescence automatic detection system, which was verified by practical cases. Results in 154 unrelated Han women and 65 male unrelated individuals of Henan Han nationality, 4 loci and 12 alleles were detected in three X-STR loci of GATA165B12 DXS6800GTA31E08, respectively. The observed values of heterozygosity were 0.6170.266 and 0.766.154 unrelated individuals of Han nationality in Henan Province, respectively. DP values were 0.806, 0.46 and 0.906, respectively. The PIC values were 0.570.28 and 0.72PE, respectively. The cumulative personal recognition ability of the three loci was 0.99015, and the cumulative non-paternal rank of the three loci was 0.99015. The probability of division is 0.6984; after chi-square test, The Hardy-Weinberg equilibrium probabilities of the three STR loci were higher than 0.05.65 male unrelated individuals of Henan Han nationality, and the haplotypes GD was 0.9529. Conclusion the three X-STR loci were studied for their species-specificity, tissue specificity, sensitivity and application in practical cases. The results showed that the three X-STR loci had better species-specificity, tissue specificity and sensitivity. The fluorescent labeling X-STR multiplex amplification test technique has good effect in practical cases.
【學(xué)位授予單位】:鄭州大學(xué)
【學(xué)位級別】:碩士
【學(xué)位授予年份】:2010
【分類號】:D919
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