Hinrichs John W J, van Blokland W T Marja, Moons Michiel J, Radersma Remco D, Radersma-van Loon Joyce H, de Voijs Carmen M A, Rappel Sophie B, Koudijs Marco J, Besselink Nicolle J M, Willems Stefan M, de Weger Roel A
From the Departments of Pathology and
From the Departments of Pathology and.
Am J Clin Pathol. 2015 Apr;143(4):573-8. doi: 10.1309/AJCP40XETVYAMJPY.
To compare next-generation sequencing (NGS) platforms with mutation-specific analysis platforms in a clinical setting, in terms of sensitivity, mutation specificity, costs, capacity, and ease of use.
We analyzed 25 formalin-fixed, paraffin-embedded lung cancer samples of different size and tumor percentage for known KRAS and EGFR hotspot mutations with two dedicated genotyping platforms (cobas [Roche Diagnostics, Almere, The Netherlands] and Rotor-Gene [QIAGEN, Venlo, The Netherlands]) and two NGS platforms (454 Genome Sequencer [GS] junior [Roche Diagnostics] and Ion Torrent Personal Genome Machine [Life Technologies, Bleiswijk, The Netherlands]).
All platforms, except the 454 GS junior, detected the mutations originally detected by Sanger sequencing and high-resolution melting prescreening and detected an additional KRAS mutation. The dedicated genotyping platforms outperformed the NGS platforms in speed and ease of use. The large sequencing capacity of the NGS platforms enabled them to deliver all mutation information for all samples at once.
Sensitivity for detecting mutations was highly comparable among all platforms. The choice for either a dedicated genotyping platform or an NGS platform is basically a trade-off between speed and genetic information.
在临床环境中，就灵敏度、突变特异性、成本、通量和易用性方面，比较新一代测序（NGS）平台与突变特异性分析平台。
我们使用两个专用基因分型平台（cobas [罗氏诊断，荷兰阿尔默勒] 和 Rotor-Gene [QIAGEN，荷兰芬洛]）以及两个 NGS 平台（454 基因组测序仪 [GS] junior [罗氏诊断] 和 Ion Torrent 个人基因组测序仪 [生命技术公司，荷兰布莱斯维克]），分析了 25 个不同大小和肿瘤百分比的福尔马林固定、石蜡包埋肺癌样本，以检测已知的 KRAS 和 EGFR 热点突变。
除 454 GS junior 外，所有平台均检测到最初通过桑格测序和高分辨率熔解预筛选检测到的突变，并检测到另外一个 KRAS 突变。专用基因分型平台在速度和易用性方面优于 NGS 平台。NGS 平台的大通量测序能力使它们能够一次性提供所有样本的所有突变信息。
所有平台检测突变的灵敏度高度可比。选择专用基因分型平台还是 NGS 平台基本上是在速度和遗传信息之间进行权衡。
