Amemiya Kenji, Hirotsu Yosuke, Oyama Toshio, Omata Masao
Genome Analysis Center, Yamanashi Central Hospital; Pathology Division, Laboratory Department, Yamanashi Central Hospital.
Genome Analysis Center, Yamanashi Central Hospital;
J Vis Exp. 2018 Mar 21(133):56943. doi: 10.3791/56943.
It is critical to determine the mutational status in cancer before administration and treatment of specific molecular targeted drugs for cancer patients. In the clinical setting, formalin-fixed paraffin-embedded (FFPE) tissues are widely used for genetic testing. However, FFPE DNA is generally damaged and fragmented during the fixation process with formalin. Therefore, FFPE DNA is sometimes not adequate for genetic testing because of low quality and quantity of DNA. Here we present a method of touch imprint cytology (TIC) to obtain genomic DNA from cancer cells, which can be observed under a microscope. Cell morphology and cancer cell numbers can be evaluated using TIC specimens. Furthermore, the extraction of genomic DNA from TIC samples can be completed within two days. The total amount and quality of TIC DNA obtained using this method was higher than that of FFPE DNA. This rapid and simple method allows researchers to obtain high-quality DNA for genetic testing (e.g., next generation sequencing analysis, digital PCR, and quantitative real time PCR) and to shorten the turnaround time for reporting results.
在给癌症患者施用和治疗特定分子靶向药物之前，确定癌症的突变状态至关重要。在临床环境中，福尔马林固定石蜡包埋（FFPE）组织被广泛用于基因检测。然而，FFPE DNA在福尔马林固定过程中通常会受损并碎片化。因此，由于DNA质量和数量较低，FFPE DNA有时不足以进行基因检测。在此，我们介绍一种触摸印记细胞学（TIC）方法，用于从癌细胞中获取基因组DNA，该DNA可在显微镜下观察到。使用TIC标本可以评估细胞形态和癌细胞数量。此外，从TIC样本中提取基因组DNA可在两天内完成。使用该方法获得的TIC DNA的总量和质量高于FFPE DNA。这种快速简单的方法使研究人员能够获得用于基因检测（如下一代测序分析、数字PCR和定量实时PCR）的高质量DNA，并缩短报告结果的周转时间。
