An integrated microfluidic system for cholangiocarcinoma diagnosis from bile by using specific affinity probes

Cancer can destroy human lives, pose significant financial burdens, and be difficult to diagnose in its early stages. For instance, the conventional antibody-body tests, such as carbohydrate antigen 19–9 (CA19–9) and carcinoembryonic antigen (CEA) which have been routinely used in laboratory tests, are not specific enough for bile duct cholangiocarcinoma (CCA), meaning that it typically remains undiagnosed until advanced stages. Thus, we aim to develop an automated integrated microfluidic system to facilitate the process of CCA diagnosis. In this system, tumor cells were automatically detected in small volume of CCA patient bile (3 mL) via three CCA-specific affinity reagents, including an antibody, an aptamer, and a glycosaminoglycan, which were mobilized on magnetic beads. This integrated microfluidic system automated the entire procedure involved bile centrifugation and CCA capture/staining in only 80 min (20, 30, & 30 min for sample treatment, cell capture, & immunostaining, respectively). Single-blind tests of clinical samples from CCA patients and non-CCA individuals with all three affinity reagents were conducted. Three out of three positive and four out of four negative samples were tested accurately, as determined through use of CCA and non-CCA patient samples. This integrated microfluidic system may therefore serve as a promising tool for improving detection of CCA.