Kärcher Janik, Schulze Britta, Dörr Aaron, Tierling Sascha, Walter Jörn
Robert Bosch GmbH, Corporate Research, Robert Bosch Campus 1, 71272 Renninge, Germany.
University of Saarland, Institute for Genetics and Epigenetics, Campus Saarbrücken, 66123 Saarbrücken, Germany.
Biomicrofluidics. 2022 Dec 6;16(6):064102. doi: 10.1063/5.0108374. eCollection 2022 Dec.
Changes in the DNA methylation landscape are associated with many diseases like cancer. Therefore, DNA methylation analysis is of great interest for molecular diagnostics and can be applied, e.g., for minimally invasive diagnostics in liquid biopsy samples like blood plasma. Sensitive detection of local methylation, which occurs in various cancer types, can be achieved with quantitative HeavyMethyl-PCR using oligonucleotides that block the amplification of unmethylated DNA. A transfer of these quantitative PCRs (qPCRs) into point-of-care (PoC) devices like microfluidic Lab-on-Chip (LoC) cartridges can be challenging as LoC systems show significantly different thermal properties than qPCR cyclers. We demonstrate how an adequate thermal model of the specific LoC system can help us to identify a suitable thermal profile, even for complex HeavyMethyl qPCRs, with reduced experimental effort. Using a simulation-based approach, we demonstrate a proof-of-principle for the successful LoC transfer of colorectal /-qPCR from Epi Procolon® colorectal carcinoma test, by avoidance of oligonucleotide interactions.
DNA甲基化格局的变化与许多疾病（如癌症）相关。因此，DNA甲基化分析在分子诊断中备受关注，并且可应用于例如血浆等液体活检样本的微创诊断。使用能阻断未甲基化DNA扩增的寡核苷酸进行定量重甲基化PCR，可以实现对各种癌症类型中发生的局部甲基化的灵敏检测。将这些定量PCR（qPCR）转移到诸如微流控芯片实验室（LoC） cartridges等即时检测（PoC）设备中可能具有挑战性，因为LoC系统的热特性与qPCR循环仪有显著差异。我们展示了特定LoC系统的适当热模型如何帮助我们识别合适的热曲线，即使对于复杂的重甲基化qPCR也是如此，同时减少实验工作量。通过基于模拟的方法，我们通过避免寡核苷酸相互作用，证明了Epi Procolon® 结直肠癌检测中的结直肠癌 /-qPCR成功转移到LoC的原理验证。
