Centre for Bio-Inspired Technology, Department of Electrical and Electronic Engineering, Imperial College London, LondonSW7 2AZ, U.K.
Imperial Centre for Translational and Experimental Medicine, Department of Surgery and Cancer, Imperial College London, LondonW12 0NN, U.K.
ACS Sens. 2022 Nov 25;7(11):3389-3398. doi: 10.1021/acssensors.2c01463. Epub 2022 Nov 11.
Prostate cancer (PCa) is the second most common cause of male cancer-related death worldwide. The gold standard of treatment for advanced PCa is androgen deprivation therapy (ADT). However, eventual failure of ADT is common and leads to lethal metastatic castration-resistant PCa. As such, the detection of relevant biomarkers in the blood for drug resistance in metastatic castration-resistant PCa patients could lead to personalized treatment options. mRNA detection is often limited by the low specificity of qPCR assays which are restricted to specialized laboratories. Here, we present a novel reverse-transcription loop-mediated isothermal amplification assay and have demonstrated its capability for sensitive detection of AR-V7 and YAP1 RNA (3 × 10 RNA copies per reaction). This work presents a foundation for the detection of circulating mRNA in PCa on a non-invasive lab-on-chip device for use at the point-of-care. This technique was implemented onto a lab-on-chip platform integrating an array of chemical sensors (ion-sensitive field-effect transistors) for real-time detection of RNA. Detection of RNA presence was achieved through the translation of chemical signals into electrical readouts. Validation of this technique was conducted with rapid detection (<15 min) of extracted RNA from prostate cancer cell lines 22Rv1s and DU145s.
前列腺癌（PCa）是全球男性癌症相关死亡的第二大常见原因。晚期 PCa 的治疗金标准是雄激素剥夺疗法（ADT）。然而，ADT 的最终失败很常见，导致致命的转移性去势抵抗性 PCa。因此，在转移性去势抵抗性 PCa 患者的血液中检测相关生物标志物以检测耐药性可能会导致个性化的治疗选择。mRNA 检测通常受到 qPCR 检测的低特异性限制，qPCR 检测仅限于专门的实验室。在这里，我们提出了一种新型的逆转录环介导等温扩增检测方法，并证明了其对 AR-V7 和 YAP1 RNA（每个反应 3×10 RNA 拷贝）的灵敏检测能力。这项工作为在即时护理点使用非侵入性片上实验室设备检测 PCa 中的循环 mRNA 奠定了基础。该技术已集成到一个片上实验室平台上，该平台集成了一系列化学传感器（离子敏感场效应晶体管），用于实时检测 RNA。通过将化学信号转化为电读数来实现 RNA 存在的检测。该技术的验证是通过快速（<15 分钟）检测从前列腺癌细胞系 22Rv1s 和 DU145s 中提取的 RNA 来进行的。
