The Integrity of Deoxyribonucleic Acid (DNA) Extracted from Whole Teeth Samples Burnt with Different Accelerants Using Two Extraction Protocols for Forensic Sex Determination

O. Iroanya, Abdul-Warith Akinshipo, J. P. Mairiga
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Abstract

Sex determination is usually the first step in forensic identification of victim(s) in disasters, jungle justice and arsonscenarios for many medical and legal reasons. The use of DNA in forensic analysis offers a good method in sexdetermination and the quality of extracted DNA is very important for downstream PCR. The aim of this study is toinvestigate the viability of DNA obtained from burnt teeth for forensic sex-determination. Two DNA extractionmethods consisting of silica based commercial kit and phenol-chloroform organic method, followed by polymerasechain amplification of amelogenin gene for sex determination were employed. Based on the DNA yield and opticaldensity values, the kit extraction method performed better than the phenol-chloroform method with 100% and85.71% success respectively. A Mann-Whitney U test of 260/280 absorbance showed no statistically significantdifference in the median absorbance for aviation fuel (median = 1.34) and gasoline (median =1.32) burnt samples.Amplification of AMEL genes using the commercial kit and phenol-chloroform method were 52.38% and 22.22%positive respectively compared to pre-extraction sex determination. This study shows that the use of the silica basedcommercial kit technique yielded higher DNA quality and quantity from whole teeth burnt with gasoline andaviation fuel as accelerants for downstream PCR amplification of AMEL genes compared to organic phenolchloroform method.
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用两种提取方法从不同促进剂烧伤的全牙样本中提取脱氧核糖核酸(DNA)的完整性,用于法医性别鉴定
由于许多医疗和法律原因,性别确定通常是灾害、丛林司法和纵火案件中受害者法医鉴定的第一步。DNA在法医分析中的应用为鉴定性别提供了一种良好的方法,提取DNA的质量对下游PCR至关重要。本研究的目的是研究从烧伤牙齿中获得的DNA用于法医性别鉴定的可行性。采用二氧化硅基商业试剂盒和苯酚-氯仿有机法两种DNA提取方法,然后进行淀粉原蛋白基因聚合酶链扩增进行性别鉴定。从DNA得率和光密度值两方面分析,试剂盒提取方法的提取率分别为100%和85.71%,优于苯酚-氯仿法。260/280吸光度的Mann-Whitney U检验显示,航空燃料(中位数= 1.34)和汽油(中位数=1.32)燃烧样品的中位数吸光度差异无统计学意义。与提取前性别鉴定相比,商用试剂盒和苯酚-氯仿法AMEL基因扩增阳性率分别为52.38%和22.22%。本研究表明,与有机酚氯仿法相比,使用硅基商业试剂盒技术可以从汽油和航空燃料作为促进剂的全牙中获得更高的DNA质量和数量,用于AMEL基因的下游PCR扩增。
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