Institute of Neurological Sciences, Italian National Research Council, Via Paolo Gaifami 18, 95126 Catania, Italy.
STMicroelectronics, Stradale Primosole 50, 95121 Catania, Italy.
Sensors (Basel). 2018 Jan 5;18(1):131. doi: 10.3390/s18010131.
The KRAS oncogene is involved in the pathogenesis of several types of cancer, particularly colorectal cancer (CRC). The most frequent mutations in this gene are associated with poor survival, increased tumor aggressiveness and resistance to therapy with anti-epidermal growth factor receptor (EGFR) antibodies. For this reason, KRAS mutation testing has become increasingly common in clinical practice for personalized cancer treatments of CRC patients. Detection methods for KRAS mutations are currently expensive, laborious, time-consuming and often lack of diagnostic sensitivity and specificity. In this study, we describe the development of a Lab-on-Chip assay for genotyping of KRAS mutational status. This assay, based on the In-Check platform, integrates microfluidic handling, a multiplex polymerase chain reaction (PCR) and a low-density microarray. This integrated sample-to-result system enables the detection of KRAS point mutations, including those occurring in codons 12 and 13 of exon 2, 59 and 61 of exon 3, 117 and 146 of exon 4. Thanks to its miniaturization, automation, rapid analysis, minimal risk of sample contamination, increased accuracy and reproducibility of results, this Lab-on-Chip platform may offer immediate opportunities to simplify KRAS genotyping into clinical routine.
KRAS 癌基因参与多种癌症的发病机制，特别是结直肠癌（CRC）。该基因的最常见突变与生存不良、肿瘤侵袭性增加以及对表皮生长因子受体（EGFR）抗体治疗的耐药性有关。因此，KRAS 基因突变检测在 CRC 患者的癌症个体化治疗中越来越普遍。目前，KRAS 突变检测方法昂贵、费力、耗时，并且常常缺乏诊断灵敏度和特异性。在本研究中，我们描述了一种用于 KRAS 突变状态基因分型的片上实验室（Lab-on-Chip）检测方法。该检测方法基于 In-Check 平台，集成了微流控处理、多重聚合酶链反应（PCR）和低密度微阵列。这种集成的样本到结果系统能够检测 KRAS 点突变，包括发生在外显子 2 的密码子 12 和 13、外显子 3 的 59 和 61、外显子 4 的 117 和 146。由于其小型化、自动化、快速分析、最小的样品污染风险、结果的准确性和重现性提高，这个片上实验室平台可能会为简化 KRAS 基因分型进入临床常规提供即时机会。
