Ohio Attorney General's Center for The Future of Forensic Science, Bowling Green State University, Bowling Green, OH 43403, USA.
Ohio Attorney General's Center for The Future of Forensic Science, Bowling Green State University, Bowling Green, OH 43403, USA; Bode Technology, 10430 Furnace Road, Suite 107, Lorton, VA 22079, USA.
Forensic Sci Int. 2022 Nov;340:111448. doi: 10.1016/j.forsciint.2022.111448. Epub 2022 Sep 5.
Heightened sensitivity of forensic DNA techniques has led to an increased variety of samples tested, often yielding complex DNA mixtures, in turn making the interpretation of profiling results more complicated. Currently, there is no prescribed upstream laboratory method to separate complex DNA mixtures by their contributors; therefore, a method is needed that could reduce mixtures into their component parts. Various cell sorting applications have the potential to be this method, if intact cells can be reliably obtained from forensic samples. Here, the effects of elution buffer and swab substrate on the recovery of intact, human, white blood cells from dried blood samples were evaluated. Approximately 328,000 cells per swab were deposited onto cotton, flocked, and dissolvable swabs. The whole-cell elution of the dried samples was evaluated with water, phosphate buffered saline, and AutoMACS® elution buffers. We demonstrate that AutoMACS® buffer is superior for the elution of intact cells, compared to phosphate buffered saline and water. When swab type was considered, the highest yield of intact cells resulted from flocked swabs, as opposed to cotton or dissolvable swabs.
法医 DNA 技术的灵敏度提高导致了更多种类的样本被检测,这些样本往往产生复杂的 DNA 混合物,进而使得分析结果更加复杂。目前,没有规定的上游实验室方法可以按供体分离复杂的 DNA 混合物;因此,需要有一种方法可以将混合物分解成其组成部分。如果可以从法医样本中可靠地获得完整细胞,那么各种细胞分选应用都有可能成为这种方法。在这里,评估了洗脱缓冲液和拭子基底对从干燥血斑样本中回收完整人类白细胞的影响。大约 328,000 个细胞/拭子被沉积在棉、植绒和可溶解拭子上。使用水、磷酸盐缓冲盐水和 AutoMACS®洗脱缓冲液对干燥样品的全细胞洗脱进行了评估。我们证明,与磷酸盐缓冲盐水和水相比,AutoMACS®缓冲液更有利于完整细胞的洗脱。当考虑拭子类型时,与棉拭子或可溶解拭子相比,植绒拭子产生的完整细胞产量最高。