Zhu Wenjie, Qian Hong, Cao Shengya, Xia Wei, Wang Xilong, Jin Jing, Wang Xin, Zhang Hao, Liu Dongsheng, Chen Ying
School of Medical Technology, Xuzhou Medical University, Xuzhou, 221004, China.
School of Medical Technology, Xuzhou Medical University, Xuzhou, 221004, China; Department of Laboratory Medicine, The Affiliated Suqian Hospital of Xuzhou Medical University, Suqian, 223800, China.
Anal Chim Acta. 2025 Jan 22;1336:343533. doi: 10.1016/j.aca.2024.343533. Epub 2024 Dec 7.
The early detection of Hepatocellular Carcinoma (HCC) is crucial for improving patient survival rates.Early diagnosis of HCC can significantly enhance treatment outcomes and reduce disease progression. Antigen detection of tumor markers is one of the important diagnostic methods for HCC. However, Traditional antigen detection methods often rely on heavy detection equipment, involve lengthy turnaround times, and must be conducted in laboratory settings. Therefore, there is a clear need for a portable, low-skill, rapid sample-to-result detection method for early HCC biomarkers.
We propose a new platform based on electrowetting-on-dielectric digital microfluidic(EWOD-DMF) for the detection of early-stage HCC biomarkers, enabling the quantitative measurement of Alpha-Fetoprotein (AFP), the proportion of AFP-L3 in total AFP (AFP-L3%), and Des-Gamma-Carboxy Prothrombin (DCP). First, serum samples are processed through the microfluidic system, achieving the separation of AFP-L3 within 10 min. Next, immunoassays are performed within 15 min, using magnetic particles to capture biomarkers such as AFP, AFP-L3, and DCP, followed by enzymatic reactions that generate detectable signals. Each chip can simultaneously detect three biomarkers from five different samples, allowing for a total of fifteen targets to be tested, with only approximately 2.4 μL of serum required for each biomarker detection. Ultimately, data are analyzed with dedicated software to quantitatively measure the HCC biomarkers. The detection limits for AFP or AFP-L3 and for DCP are 0.24 ng/mL and 1.89 ng/mL, respectively.
This study presents a EWOD-DMF platform for early-stage HCC diagnosis, capable of simultaneously detecting multiple samples and biomarkers, thus improving detection efficiency and diagnostic accuracy. Moreover, the platform has POCT capability, with advantages in portability and cost-effectiveness, providing clinicians and primary healthcare institutions with a fast and convenient solution for early-stage HCC diagnosis.
肝细胞癌（HCC）的早期检测对于提高患者生存率至关重要。HCC的早期诊断可显著改善治疗效果并减缓疾病进展。肿瘤标志物的抗原检测是HCC重要的诊断方法之一。然而，传统的抗原检测方法通常依赖笨重的检测设备，周转时间长，且必须在实验室环境中进行。因此，迫切需要一种用于早期HCC生物标志物的便携式、低技术要求、快速的样本到结果检测方法。
我们提出了一种基于介电电泳数字微流控（EWOD-DMF）的新平台，用于检测早期HCC生物标志物，能够定量检测甲胎蛋白（AFP）、AFP-L3在总AFP中的比例（AFP-L3%）以及异常凝血酶原（DCP）。首先，血清样本通过微流控系统进行处理，在10分钟内实现AFP-L3的分离。接下来，在15分钟内进行免疫测定，使用磁性颗粒捕获AFP、AFP-L3和DCP等生物标志物，随后进行酶促反应产生可检测信号。每个芯片可同时检测来自五个不同样本的三种生物标志物，总共可测试十五个靶点，每个生物标志物检测仅需约2.4μL血清。最终，使用专用软件分析数据以定量测量HCC生物标志物。AFP或AFP-L3以及DCP的检测限分别为0.24 ng/mL和1.89 ng/mL。
本研究提出了一种用于早期HCC诊断的EWOD-DMF平台，能够同时检测多个样本和生物标志物，从而提高检测效率和诊断准确性。此外，该平台具有即时检验（POCT）能力，在便携性和成本效益方面具有优势，为临床医生和基层医疗机构提供了一种快速便捷的早期HCC诊断解决方案。
