title: Microfluidic purification and concentration of malignant pleural effusions for improved molecular and cytomorphological diagnostics.
abstract: Evaluation of pleural fluids for metastatic cells is a key component of diagnostic cytopathology. However, a large background of smaller leukocytes and/or erythrocytes can make accurate diagnosis difficult and reduce specificity in identification of mutations of interest for targeted anti-cancer therapies. Here, we describe an automated microfluidic system (Centrifuge Chip) which employs microscale vortices for the size-based isolation and concentration of cancer cells and mesothelial cells from a background of blood cells. We are able to process non-diluted pleural fluids at 6 mL/min and enrich target cells significantly over the background; we achieved improved purity in all patient samples analyzed. The resulting isolated and viable cells are readily available for immunostaining, cytological analysis, and detection of gene mutations. To demonstrate the utility towards aiding companion diagnostics, we also show improved detection accuracy of KRAS gene mutations in lung cancer cells processed using the Centrifuge Chip, leading to an increase in the area under the curve (AUC) of the receiver operating characteristic from 0.90 to 0.99. The Centrifuge Chip allows for rapid concentration and processing of large volumes of bodily fluid samples for improved cytological diagnosis and purification of cells of interest for genetic testing, which will be helpful for enhancing diagnostic accuracy.
authors.affiliation: Department of Bioengineering, University of California Los Angeles, Los Angeles, California, United States of America.
authors.forename: James
authors.initials: J
authors.lastname: Che
authors.forename: Albert J
authors.initials: AJ
authors.lastname: Mach
authors.forename: Derek E
authors.initials: DE
authors.lastname: Go
authors.forename: Ish
authors.initials: I
authors.lastname: Talati
authors.forename: Yong
authors.initials: Y
authors.lastname: Ying
authors.forename: Jianyu
authors.initials: J
authors.lastname: Rao
authors.forename: Rajan P
authors.initials: RP
authors.lastname: Kulkarni
authors.forename: Dino
authors.initials: D
authors.lastname: Di Carlo
mesh_terms: D019540:Area Under Curve; D014408:Biomarkers, Tumor / Q000235:genetics; D004848:Epithelium / Q000473:pathology; D006801:Humans; D008175:Lung Neoplasms / Q000175:diagnosis / Q000235:genetics / Q000473:pathology; D044085:Microfluidics / Q000379:methods; D057089:Pathology, Molecular / Q000379:methods; D016066:Pleural Effusion, Malignant / Q000175:diagnosis* / Q000235:genetics / Q000473:pathology*; D011518:Proto-Oncogene Proteins / Q000235:genetics; D016283:Proto-Oncogene Proteins p21(ras); D012372:ROC Curve; D012680:Sensitivity and Specificity; D018631:ras Proteins / Q000235:genetics
chemical_list: D014408:Biomarkers, Tumor; C117307:KRAS protein, human; D011518:Proto-Oncogene Proteins; D016283:Proto-Oncogene Proteins p21(ras); D018631:ras Proteins