Opel Kerry L, Chung Denise T, Drábek Jirí, Butler John M, McCord Bruce R
International Forensic Research Institute, Florida International University, 11200 SW 8th Street, Miami, FL 33199, USA.
J Forensic Sci. 2007 Nov;52(6):1263-71. doi: 10.1111/j.1556-4029.2007.00584.x.
This paper describes a developmental validation study of three Miniplex sets covering 12 of the 13 CODIS loci. As these new sets will be used for the analysis of degraded and low level DNA, the validation studies were performed using 100-125 pg of DNA, the lowest input level at which peak balance, peak intensity, and allele consistency were stable. To demonstrate the applicability of the Miniplex sets to forensic casework, these validation studies were completed in accordance with the Scientific Working Group on DNA Analysis Methods (SWGDAM). A range of tests were performed including studies of concordance with standard multiplex kits, sensitivity and reproducibility, and PCR amplification conditions. Additionally, studies of mixtures, nonhuman and environmentally degraded DNA, and simulated forensic samples were performed. Our results demonstrate that Miniplex STR amplification procedures are a robust and sensitive tool for the analysis of degraded DNA.
本文描述了一项针对覆盖13个联合DNA索引系统(CODIS)位点中12个位点的三种复合扩增试剂盒的开发验证研究。由于这些新试剂盒将用于降解和低水平DNA的分析,验证研究使用100 - 125皮克的DNA进行,这是峰平衡、峰强度和等位基因一致性稳定的最低输入水平。为证明复合扩增试剂盒在法医案件工作中的适用性,这些验证研究是按照DNA分析方法科学工作组(SWGDAM)的要求完成的。进行了一系列测试,包括与标准复合扩增试剂盒的一致性研究、灵敏度和可重复性研究以及PCR扩增条件研究。此外,还进行了混合物、非人类和环境降解DNA以及模拟法医样本的研究。我们的结果表明,复合短串联重复序列(STR)扩增程序是分析降解DNA的一种强大且灵敏的工具。