Frégeau C J, Vanstone H, Borys S, McLean D, Maroun J A, Birnboin H C, Fourney R M
Royal Canadian Mounted Police, National DNA Data Bank, Ottawa, ON.
J Forensic Sci. 2001 Sep;46(5):1180-90.
A preliminary study was conducted to assess the capability of a new alcohol-based tissue fixative, GenoFix, to preserve DNA from biopsy tissues stored at room temperature and/or -20 degrees C in a freezer, for subsequent short tandem repeat (STR) DNA typing analysis. Fresh human smooth muscle samples were stored at room temperature in GenoFix for one month and up to one year and seven months before being processed using the megaplex STR systems, AmpFlSTR Profiler Plus and AmpFlSTR COfiler. Alternatively, muscle tissues in GenoFix were placed at -20 degrees C in a freezer for up to 3 1/2 years following two to three months in the fixative at room temperature. DNA analysis was also carried out on tissues stored in GenoFix for one month at room temperature and subsequently paraffin-embedded and stored at room temperature for four years. The AmpFlSTR Profiler Plus and AmpFlSTR COfiler STR profiles produced, using DNA extracted from all fixed tissue samples, were of very good quality. The fluorescent signals were well balanced across the nine STR loci or six loci comprised in the megaplexes surveyed and profiles showed no differences with those observed for the control blood of the respective donor patients. Continuous exposure to GenoFix at room temperature (up to one year and seven months) did not compromise the STR typing analysis of the fixed tissues. No adverse effects were noted on the STR typeability of tissues fixed with GenoFix and stored at -20 degrees C in a freezer for up to 3 1/2 years. STR profiles generated from the paraffin-embedded tissues fixed in GenoFix were of excellent quality. This preliminary study suggests that GenoFix can be used to store tissue samples at room temperature for up to one year and seven months or at -20 degrees C in a freezer for longer storage (up to 3 1/2 years). This new and odorless tissue fixative promotes tissue and DNA preservation in a very effective manner and as such may prove useful in criminal investigations or mass disaster identifications carried out in remote locations and in which a small or large number of tissue samples are collected for further analyses.
进行了一项初步研究,以评估一种新型酒精基组织固定剂GenoFix在保存室温及/或-20℃冷冻保存的活检组织中的DNA方面的能力,以便随后进行短串联重复序列(STR)DNA分型分析。新鲜人类平滑肌样本在室温下于GenoFix中保存1个月至1年零7个月,然后使用复合STR系统AmpFlSTR Profiler Plus和AmpFlSTR COfiler进行处理。另外,在室温下于固定剂中放置两到三个月后,将GenoFix中的肌肉组织置于-20℃的冰箱中保存长达3年半。还对在室温下于GenoFix中保存1个月、随后石蜡包埋并在室温下保存4年的组织进行了DNA分析。使用从所有固定组织样本中提取的DNA产生的AmpFlSTR Profiler Plus和AmpFlSTR COfiler STR图谱质量非常好。在所检测的复合体系中的9个STR位点或6个位点上,荧光信号平衡良好,图谱与相应供体患者的对照血液所观察到的图谱无差异。在室温下持续暴露于GenoFix(长达1年零7个月)不会损害固定组织的STR分型分析。对于用GenoFix固定并在-20℃的冰箱中保存长达3年半的组织的STR分型能力,未观察到不利影响。由GenoFix固定的石蜡包埋组织产生的STR图谱质量优异。这项初步研究表明,GenoFix可用于在室温下保存组织样本长达1年零7个月,或在-20℃的冰箱中进行更长时间的保存(长达3年半)。这种新型无味组织固定剂能非常有效地促进组织和DNA的保存,因此在偏远地区进行的刑事调查或大规模灾难鉴定中可能证明是有用的,在这些情况下会收集少量或大量组织样本进行进一步分析。