Prasad Elisha, Hitchcock Catherine, Raymond Jennifer, Cole Andrew, Barash Mark, McNevin Dennis, van Oorschot Roland A H
Centre for Forensic Science, School of Mathematical & Physical Sciences, Faculty of Science, University of Technology Sydney, PO Box 123, Broadway, NSW 2007, Australia.
NSW Health Pathology, Forensic & Analytical Science Service, PO Box 162, Lidcombe, NSW 1825, Australia.
Forensic Sci Int. 2022 Jan;330:111101. doi: 10.1016/j.forsciint.2021.111101. Epub 2021 Nov 10.
Over the recent few years, several DNA collection techniques and methodologies have been published for the recovery of DNA from fired cartridge cases. In this study, swabbing, the DNA collection technique currently used in our jurisdiction (NSW, Australia), was compared with tape lifting and soaking to assess DNA recovery rates, DNA quality and profile quality. Brass .22LR and 9mmP cartridges were used as they are the most commonly encountered in our jurisdiction. The cartridges (n = 107) were loaded into cleaned firearm magazines by three volunteers of unknown shedder status, to mimic routine casework sample types. Half of the handled cartridges were fired whilst the other half were kept unfired. STR genotypes were produced at both 29 and 30 PCR cycles to evaluate which improved handler allele detection. DNA recovery rates showed that swabbing recovered significantly less DNA than tape lifting and soaking. Whilst there were no significant differences between tape lifting and soaking, tape lifting, on average, yielded more DNA than soaking. The calibre of ammunition had no influence on DNA recovery and in line with expectations, firing was found to decrease DNA recovery for all three sampling techniques. Assessment of DNA quality showed no evidence of PCR inhibition in any of the samples for this study. However, degradation indices showed that most samples were slightly to moderately degraded. Fewer handler alleles were detected from both fired tape lifted and soaked cartridges than unfired cartridges. Whilst 30 amplification cycles allowed for the detection of slightly more handler alleles, no statistically significant differences were found between 29 and 30 PCR cycles. Nonetheless, 50% of the profiles from unfired soaked cartridges that were non-uploadable after 29 cycles were uploadable after 30 cycles. Furthermore, 83% of profiles from unfired cartridges that were tape lifted were uploadable onto our jurisdiction's database at both 29 and 30 PCR cycles. All magazine controls, despite cleaning, contained some level of background DNA. Furthermore, increasing the number of PCR cycles to 30 also increased the detection of non-handler alleles in DNA profiles. Our results suggest tape lifting yields more uploadable profiles from unfired and fired cartridge cases than swabbing but also more adventitious (non-handler) alleles. However additional research will be needed to evaluate the full potential of this method.
在过去几年中,已经发表了几种从已发射子弹壳中提取DNA的采集技术和方法。在本研究中,将我们辖区(澳大利亚新南威尔士州)目前使用的DNA采集技术拭子法,与胶带粘取法和浸泡法进行比较,以评估DNA回收率、DNA质量和图谱质量。使用.22LR黄铜和9mmP子弹壳,因为它们是我们辖区最常见的。107个子弹壳由三名 shedding 状态未知的志愿者装入清洁过的枪支弹匣,以模拟常规案件样本类型。一半处理过的子弹壳被发射,另一半未发射。在29个和30个PCR循环下产生STR基因型,以评估哪种方法能提高经手人等位基因检测率。DNA回收率表明,拭子法回收的DNA明显少于胶带粘取法和浸泡法。虽然胶带粘取法和浸泡法之间没有显著差异,但平均而言,胶带粘取法比浸泡法产生的DNA更多。弹药口径对DNA回收没有影响,并且正如预期的那样,发现发射会降低所有三种采样技术的DNA回收率。DNA质量评估表明,本研究的任何样本中均未发现PCR抑制的证据。然而,降解指数表明,大多数样本有轻微至中度降解。从已发射的经胶带粘取和浸泡处理的子弹壳中检测到的经手人等位基因比未发射的子弹壳少。虽然30个扩增循环能检测到稍多的经手人等位基因,但在29个和30个PCR循环之间未发现统计学上的显著差异。尽管如此,29个循环后无法上传的未发射浸泡子弹壳样本中,有50%在30个循环后可以上传。此外,在29个和30个PCR循环下,83%的经胶带粘取的未发射子弹壳样本图谱可以上传到我们辖区的数据库中。所有弹匣对照,尽管经过清洁,仍含有一定水平的背景DNA。此外,将PCR循环数增加到30也增加了DNA图谱中非经手人等位基因的检测。我们的结果表明,与拭子法相比,胶带粘取法从未发射和已发射的子弹壳中产生的可上传图谱更多,但也有更多偶然的(非经手人)等位基因。然而,需要进一步的研究来评估这种方法的全部潜力。