title: On-chip Extraction of Intracellular Molecules in White Blood Cells from Whole Blood.
abstract: The extraction of virological markers in white blood cells (WBCs) from whole blood--without reagents, electricity, or instruments--is the most important first step for diagnostic testing of infectious diseases in resource-limited settings. Here we develop an integrated microfluidic chip that continuously separates WBCs from whole blood and mechanically ruptures them to extract intracellular proteins and nucleic acids for diagnostic purposes. The integrated chip is assembled with a device that separates WBCs by using differences in blood cell size and a mechanical cell lysis chip with ultra-sharp nanoblade arrays. We demonstrate the performance of the integrated device by quantitatively analyzing the levels of extracted intracellular proteins and genomic DNAs. Our results show that compared with a conventional method, the device yields 120% higher level of total protein amount and similar levels of gDNA (90.3%). To demonstrate its clinical application to human immunodeficiency virus (HIV) diagnostics, the developed chip was used to process blood samples containing HIV-infected cells. Based on PCR results, we demonstrate that the chip can extract HIV proviral DNAs from infected cells with a population as low as 10(2)/μl. These findings suggest that the developed device has potential application in point-of-care testing for infectious diseases in developing countries.
authors.affiliation: School of Mechatronics, Gwangju Institute of Science and Technology (GIST), Gwangju, 500-712, Republic of Korea.
authors.forename: Jongchan
authors.initials: J
authors.lastname: Choi
authors.affiliation: School of Mechatronics, Gwangju Institute of Science and Technology (GIST), Gwangju, 500-712, Republic of Korea.
authors.forename: Ji-chul
authors.initials: JC
authors.lastname: Hyun
authors.affiliation: School of Mechatronics, Gwangju Institute of Science and Technology (GIST), Gwangju, 500-712, Republic of Korea.|Department of Medical System Engineering, Gwangju Institute of Science and Technology (GIST), Gwangju, 500-712, Republic of Korea.
authors.forename: Sung
authors.initials: S
authors.lastname: Yang
mesh_terms: D001769:Blood / Q000378:metabolism*; D001798:Blood Proteins / Q000737:chemistry / Q000302:isolation & purification*; D002458:Cell Fractionation / Q000379:methods*; D002478:Cells, Cultured; D004247:DNA / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification*; D006801:Humans; D056656:Lab-On-A-Chip Devices; D007962:Leukocytes / Q000737:chemistry*
chemical_list: D001798:Blood Proteins; D004247:DNA