Roche Sequencing Solutions, Pleasanton, California.
Signature Diagnostics, Potsdam, Germany.
J Mol Diagn. 2021 Apr;23(4):399-406. doi: 10.1016/j.jmoldx.2021.01.006. Epub 2021 Jan 23.
Formalin-fixed, paraffin-embedded (FFPE) tissue is the most commonly used material for tumor molecular profiling, therapy selection, and prognostication. Tumor tissue enrichment by tissue dissection is highly recommended to generate quality data reproducibly for use in downstream assays, such as real-time PCR and next-generation sequencing. The aim of this study was to evaluate the performance of the automated tissue dissection tool AVENIO Millisect System compared with a manual dissection method using 18 FFPE tissue specimens. The study assessed performance of these two methods with paraffinized and deparaffinized sections at 5- and 10-μm thickness as well as at low (5% to 10%) and high (>50%) tumor content. In addition, compatibility with various nucleic acid and protein extraction methods was assessed. Overall, dissection by Millisect resulted in statistically significantly higher yields of nucleic acids and protein compared with manual dissection (P = 0.00524). In downstream analysis on a statistically nonpowered sample set, EGFR mutation testing by PCR led to highly concordant results, and next-generation sequencing testing yielded significantly higher allelic frequencies when tissue was dissected by Millisect compared with manual scraping, demonstrating noninferiority of the automated method. In summary, the AVENIO Millisect System may replace manual labor and support automation of FFPE tumor tissue workflows in clinical molecular laboratories with high testing volumes with adequate validation.
福尔马林固定、石蜡包埋(FFPE)组织是最常用于肿瘤分子分析、治疗选择和预后的材料。强烈建议通过组织解剖来富集肿瘤组织,以生成可重复使用的高质量数据,用于下游检测,如实时 PCR 和下一代测序。本研究旨在评估自动化组织解剖工具 AVENIO Millisect 系统与手动解剖方法在 18 个 FFPE 组织标本中的性能比较。该研究评估了这两种方法在 5-μm 和 10-μm 厚的石蜡和去石蜡切片以及低(5%至 10%)和高(>50%)肿瘤含量下的性能。此外,还评估了与各种核酸和蛋白质提取方法的兼容性。总体而言,与手动解剖相比,Millisect 解剖在核酸和蛋白质方面的产量统计学上显著更高(P=0.00524)。在一个统计学上无能力的样本集的下游分析中,PCR 检测的 EGFR 突变测试结果高度一致,当使用 Millisect 进行组织解剖时,下一代测序测试的等位基因频率显著更高,表明自动化方法具有非劣效性。总之,AVENIO Millisect 系统可以替代人工劳动,并在具有高检测量的临床分子实验室中支持 FFPE 肿瘤组织工作流程的自动化,具有充分的验证。