本文選題：法醫(yī)遺傳學(xué) + miniSTR��； 參考：《四川大學(xué)》2007年碩士論文

【摘要】： 目的本課題旨在建立新的miniSTR熒光標記復(fù)合擴增體系，并按照美國DNA分析方法技術(shù)工作組(TWGDAM)的指導(dǎo)方案，進行法醫(yī)學(xué)應(yīng)用性研究。方法從已有國內(nèi)頻率資料的STR基因座中篩選出可用于設(shè)計MiniSTR引物的基因座，進行引物設(shè)計；用未標記的新引物在銀染顯色方法下進行復(fù)合擴增實驗，選出復(fù)合成功基因座定購熒光標記引物；制作等位基因分型標準物；采用美國ABI310遺傳分析儀進行熒光標記引物復(fù)合擴增試驗，并優(yōu)化擴增條件，對不同退火溫度下復(fù)合擴增的效果進行研究；對復(fù)合擴增系統(tǒng)的靈敏度、基因座的種屬特異性、案例應(yīng)用、降解模型、陳舊樣本等法醫(yī)學(xué)應(yīng)用進行研究。結(jié)果成功設(shè)計了D19S591，D2S2944，D18S872基因座的小于130bp的miniSTR擴增引物，成功建立了D19S591，D2S2944，D18S872的miniSTR熒光標記復(fù)合擴增體系。這一擴增體系條件易于優(yōu)化，采用國產(chǎn)Taq酶進行擴增，即可得到滿意的擴增產(chǎn)物和分型結(jié)果。相對于國外昂貴的試劑盒，它是一種成本低廉但效率高的STR基因座復(fù)合擴增體系。法醫(yī)學(xué)應(yīng)用性研究表明：三個基因座有較高的種屬特異性；該體系的檢測靈敏度為0．25ng模板DNA；通過6個實際檢案證明，該體系能用于法醫(yī)學(xué)個人識別和親權(quán)鑒定；通過對陳舊樣本和降解模型的研究證明，該體系對降解檢材有良好的擴增效果，較常規(guī)大片段試劑盒擴增成功率高，可用于常規(guī)試劑盒擴增失敗的降解樣本的分型。結(jié)論D19S591，D2S2944，D185872的miniSTR熒光標記復(fù)合擴增體系可應(yīng)用ABI-310檢測平臺，獲得可靠的遺傳學(xué)數(shù)據(jù)。該體系擴增條件易于優(yōu)化，成本低廉，分型結(jié)果穩(wěn)定，種屬特異性好，靈敏度和準確度高，，對降解檢材擴增成功率高，可以應(yīng)用于法醫(yī)學(xué)實際檢案，特別適合對陳舊樣本和降解檢材的法醫(yī)學(xué)檢測。
[Abstract]:Objective to establish a new miniSTR fluorescent labeling multiplex amplification system, and to carry out forensic application research according to the guidance of the DNA analysis technique working group in the United States.Methods STR loci which could be used to design MiniSTR primers were screened from STR loci with existing domestic frequency data, and the primers were designed, and the multiplex amplification experiments were carried out with new unlabeled primers by silver staining method.The fluorescent marker primers were selected for complex successful loci, the allelic genotyping standard materials were made, and the multiplex amplification test of fluorescent marker primers was carried out by ABI310 genetic analyzer in the United States, and the amplification conditions were optimized.The effects of multiplex amplification at different annealing temperatures were studied, and the sensitivity of multiplex amplification system, gene locus specificity, case application, degradation model, old samples and other forensic applications were studied.Results the miniSTR primers of D19S591mD2S2944nD18S872 locus were designed successfully, and the multiplex amplification system of D19S591D2S294nD18S872 was successfully established.The conditions of this amplification system are easy to be optimized, and satisfactory amplification products and typing results can be obtained by using domestic Taq enzyme.Compared with foreign expensive kit, it is a low cost and high efficiency STR locus multiplex amplification system.Forensic application studies showed that the three loci had high species-specificity, the sensitivity of the system was 0.25ng template DNA, and the system could be used for forensic personal identification and paternity identification.The study on the old samples and degradation models showed that the system had a good amplification effect on the degradation samples, and had a higher success rate than the conventional large fragment kits, and could be used for the typing of degraded samples which failed in the amplification of the conventional kits.Conclusion the miniSTR multiplex amplification system of D19S591D2S2944 and D185872 can be used for ABI-310 detection to obtain reliable genetic data.The system is easy to optimize, low cost, stable typing results, good species specificity, high sensitivity and accuracy, high success rate of amplification of degradable samples, and can be used in forensic medical practice.It is especially suitable for forensic examination of old samples and biodegradable 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

【參考文獻】

相關(guān)期刊論文 前1條

1 李英碧,吳謹,侯一平,張霽,廖淼,張林,陳國弟;STR基因座熒光標記復(fù)合擴增檢測及其法醫(yī)學(xué)應(yīng)用[J];法醫(yī)學(xué)雜志;2005年02期

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