Huijsmans Ronald, Damen Jan, van der Linden Hans, Hermans Mirjam
Multidisciplinary Laboratory of Molecular Diagnostics, Jeroen Bosch Hospital, 's-Hertogenbosch, The Netherlands.
J Mol Diagn. 2007 Apr;9(2):205-13. doi: 10.2353/jmoldx.2007.060059.
To identify issues of sample mix-ups, various molecular techniques are currently used. These techniques, however, are time consuming and require experience and/or DNA sequencing equipment or have a relatively high risk of errors because of contamination. Therefore, a quick and straightforward single nucleotide polymorphism (SNP) profiling assay was developed to link human tissues to a source. SNPs are common sequence variations in the human genome, and each individual has a unique combination of these nucleotide variations. Using potentially mislabeled paraffin-embedded tissues, DNA was extracted and SNP profiles were determined by real-time polymerase chain reaction analysis of the purified DNA using a selection of 10 commercially available SNP amplification assays. These profiles were compared with profiles of the supposed owners. All issues (34 in total) of potential sample mix-ups during the last 3 years were adequately solved, with six cases described here. The SNP profiling assay provides a quick (within 24 hours), easy, and reliable way to link human samples to a source, without polymerase chain reaction postprocessing. The chance for two randomly chosen individuals to have an identical profile is 1 in 18,000. Solving potential sample mix-ups will secure downstream evaluations and critical decisions concerning the patients involved.
为了识别样本混淆问题,目前使用了各种分子技术。然而,这些技术耗时且需要经验和/或DNA测序设备,或者由于污染而存在相对较高的错误风险。因此,开发了一种快速且直接的单核苷酸多态性(SNP)分析方法,用于将人体组织与来源进行关联。SNP是人类基因组中常见的序列变异,每个人都有这些核苷酸变异的独特组合。使用可能标记错误的石蜡包埋组织,提取DNA,并通过使用10种市售SNP扩增检测试剂盒对纯化后的DNA进行实时聚合酶链反应分析来确定SNP图谱。将这些图谱与假定所有者的图谱进行比较。过去3年中所有潜在样本混淆问题(总共34个)均得到妥善解决,本文描述了其中6个案例。SNP分析方法提供了一种快速(24小时内)、简便且可靠的方法,可将人体样本与来源进行关联,无需进行聚合酶链反应后处理。两个随机选择的个体拥有相同图谱的概率为1/18000。解决潜在的样本混淆问题将确保对相关患者进行下游评估和关键决策。