Department of Clinical Laboratory, The Second Hospital of Shandong University, Jinan, Shandong, China.
Key Laboratory for Liquid-Solid Structural Evolution and Processing of Materials, Ministry of Education, Shandong University, Jinan, Shandong, China.
Acta Biomater. 2022 Sep 1;149:287-296. doi: 10.1016/j.actbio.2022.06.037. Epub 2022 Jun 25.
piR-31,143 has been identified as a potential biomarker for the diagnosis of colorectal cancer (CRC). However, the current detection methods have complicated operations and high cost, which restrict its clinical application. In the present work, we reported a new photoelectrochemical (PEC) biosensor based on MoS@ReS/TiC hybrid and duplex-specific nuclease (DSN) assisted signal amplification mechanism for ultrasensitive detection of piRNA-31,143 from human serum. The formation of the type I heterostructure between MoS and ReS and the doping of TiC contributed to high photocurrent response. The presence of piR-31,143 triggered the chain displacement reaction and enzymatic cyclic amplification reaction on the electrode surface with the assistance of DSN, leading to the collapse of the composite probe system. Consequently, the photocurrent of the PEC biosensor was proportional to the concentration of piR-31,143. The linear detection range and calculated detection limit of the PEC biosensor were 10-10 fM and 23 aM, respectively. The stability of the photocurrent under 15 consecutive on-off irradiations (with a relative standard deviation of 1.17%) and the specific response to piR-31,143 demonstrated the reliability of the PEC biosensor. In addition, the practicability of the PEC biosensor was verified by batch detection of human serum. The area under the receiver operating characteristic curve used to distinguish CRC patients from healthy controls was 0.942 with 100% specificity, demonstrating the developed method is a promising approach for the diagnosis of CRC. STATEMENT OF SIGNIFICANCE: The clinical translation of piRNAs for cancer diagnosis is hindered by efficacy of detection techniques due to tedious sample processing and costly instrumentation. Herein, we fabricated a photoelectrochemical biosensor for the ultrasensitive detection of piR-31,143 with 10 μL serum in vitro. MoS@ReS/TiC greatly enhances the photocurrent response while duplex-specific nuclease improves the detection sensitivity and avoids false positives. By transforming the recognition sequence of the probe, the sensor can be applied to a variety of piRNAs detection for different diseases. In addition, the electrode can be recycled which is beneficial to reduce the cost of detection. With suitable automation and further optimization, our work may serve as core component in the development of an accurate and efficient diagnosis method.
piR-31,143 已被鉴定为结直肠癌（CRC）诊断的潜在生物标志物。然而，目前的检测方法操作复杂且成本高，限制了其临床应用。在本工作中，我们报道了一种基于 MoS@ReS/TiC 杂化和双链特异性核酸酶（DSN）辅助信号放大机制的新型光电化学（PEC）生物传感器，用于超灵敏检测人血清中的 piRNA-31,143。MoS 和 ReS 之间形成 I 型异质结，TiC 的掺杂有助于提高光电流响应。在 DSN 的辅助下，piR-31,143 的存在引发了电极表面上的链置换反应和酶循环扩增反应，导致复合探针系统的崩溃。因此，PEC 生物传感器的光电流与 piR-31,143 的浓度成正比。PEC 生物传感器的线性检测范围和计算检测限分别为 10-10 fM 和 23 aM。在 15 次连续开-关照射下（相对标准偏差为 1.17%）光电流的稳定性和对 piR-31,143 的特异性响应证明了 PEC 生物传感器的可靠性。此外，通过批量检测人血清验证了 PEC 生物传感器的实用性。用于区分 CRC 患者和健康对照者的受试者工作特征曲线下面积为 0.942，特异性为 100%，表明所开发的方法是 CRC 诊断的一种有前途的方法。
由于检测技术的繁琐样品处理和昂贵的仪器，piRNAs 用于癌症诊断的临床转化受到限制。在此，我们体外构建了一种用于超灵敏检测 piR-31,143 的光电化学生物传感器，仅需 10 μL 血清。MoS@ReS/TiC 极大地增强了光电流响应，而双链特异性核酸酶提高了检测灵敏度并避免了假阳性。通过改变探针的识别序列，该传感器可应用于多种 piRNAs 检测不同疾病。此外，电极可以回收，有利于降低检测成本。通过适当的自动化和进一步优化，我们的工作可能成为开发准确有效的诊断方法的核心组件。
