Nicklas Janice A, Buel Eric
Vermont Forensic Laboratory, Department of Public Safety, 103 S. Main St., Waterbury, VT 05671, USA.
J Forensic Sci. 2006 Sep;51(5):1005-15. doi: 10.1111/j.1556-4029.2006.00211.x.
A single duplex assay to determine both the amount of total human DNA and the amount of male DNA in a forensic sample has been developed. This assay is based on TaqMan technology and uses the multicopy Alu sequence to quantitate total human DNA and the multicopy DYZ5 sequence to quantitate Y chromosomal (male) DNA. The assay accepts a wide concentration range of input DNA (2 muL of 64 ng/microL to 0.5 pg/microL), and also allows detection of PCR failure. The PCR product sizes Alu (127 bp) and DYZ5 (137bp) approximate that of the smaller short tandem repeats (STRs) which should make the assay predictive of STR success with degraded DNA. The assay was optimized for probe/primer concentrations and BSA addition and validated on its reproducibility, on its human specificity, on its nonethnic variability, for artificial mixtures and adjudicated casework, for the effect of inhibitors and for state of DNA degradation. This assay should prove very usual in forensic analyses because knowing the relative amounts of male versus female DNA can allow the examiner to decide which samples may yield the most probative value in a case or direct the samples to methods that would yield the greatest information.
已开发出一种单一的双重检测方法，用于确定法医样本中人类总DNA的量和男性DNA的量。该检测方法基于TaqMan技术，使用多拷贝Alu序列定量人类总DNA，使用多拷贝DYZ5序列定量Y染色体（男性）DNA。该检测方法可接受宽浓度范围的输入DNA（64 ng/μL的2 μL至0.5 pg/μL），还能检测PCR失败情况。Alu（127 bp）和DYZ5（137 bp）的PCR产物大小与较小的短串联重复序列（STR）相近，这应使该检测方法能够预测降解DNA的STR成功情况。该检测方法针对探针/引物浓度和添加牛血清白蛋白进行了优化，并在其重现性、人类特异性、非种族变异性、人工混合物和已裁决案件、抑制剂的影响以及DNA降解状态方面进行了验证。该检测方法在法医分析中应证明非常有用，因为了解男性与女性DNA的相对量可以让检验人员决定哪些样本在案件中可能产生最大的证明价值，或者将样本导向能产生最大信息量的方法。
