title: On-chip purification and detection of hepatitis C virus RNA from human plasma.
abstract: Hepatitis C virus (HCV) is one of the main causes of chronic liver disease worldwide. The diagnosis and monitoring of HCV infection is a crucial need in the clinical management. The conventional diagnostic technologies are challenged when trying to address molecular diagnostics, especially because they require a complex and time-consuming sample preparation phase. Here, a new concept based on surface functionalization was applied to viral RNA purification: first of all polydimethylsiloxane (PDMS) flat surfaces were modified to hold RNA adsorption. After a careful chemical and morphological analysis of the modified surfaces, the functionalization protocols giving the best RNA adsorbing surfaces were applied to PDMS microdevices. The functionalized microdevices were then used for RNA purification from HCV infected human plasma samples. RNA purification and RT were successfully performed in the same microdevice chamber, saving time of analysis, reagents, and labor. The PCR protocol for HCV cDNA amplification was also implemented in the microdevice, demonstrating that the entire process of HCV analysis, from plasma to molecular readout, could be performed on-chip. Not only HCV but also other microdevice-based viral RNA detection could therefore result in a successful Point-of-Care (POC) diagnostics for resource-limited settings.
authors.affiliation: Fondazione Bruno Kessler, Laboratory of Biomolecular Sequence and Structure Analysis for Health, Via Sommarive 18, I-38123, Povo, Trento, Italy.
authors.forename: V
authors.initials: V
authors.lastname: Vaghi
authors.affiliation: Fondazione Bruno Kessler, Laboratory of Biomolecular Sequence and Structure Analysis for Health, Via Sommarive 18, I-38123, Povo, Trento, Italy; CNR - Consiglio Nazionale delle Ricerche, Istituto di Biofisica, Via alla Cascata 56/C, I-38123, Povo, Trento, Italy. Electronic address: cpotrich@fbk.eu.
authors.forename: C
authors.initials: C
authors.lastname: Potrich
authors.affiliation: Fondazione Bruno Kessler, Laboratory of Biomolecular Sequence and Structure Analysis for Health, Via Sommarive 18, I-38123, Povo, Trento, Italy.
authors.forename: L
authors.initials: L
authors.lastname: Pasquardini
authors.affiliation: Fondazione Bruno Kessler, Laboratory of Biomolecular Sequence and Structure Analysis for Health, Via Sommarive 18, I-38123, Povo, Trento, Italy; CNR - Consiglio Nazionale delle Ricerche, Istituto di Biofisica, Via alla Cascata 56/C, I-38123, Povo, Trento, Italy.
authors.forename: L
authors.initials: L
authors.lastname: Lunelli
authors.affiliation: Fondazione Bruno Kessler, Center for Materials and Microsystem (CMM), Via Sommarive 18, I-38123, Povo, Trento, Italy.
authors.forename: L
authors.initials: L
authors.lastname: Vanzetti
authors.affiliation: Department of Biomedical and Clinical Sciences "Luigi Sacco", Section of Infectious Diseases and Immunopathology, University of Milan, Milano, Italy.
authors.forename: E
authors.initials: E
authors.lastname: Ebranati
authors.affiliation: Department of Biomedical and Clinical Sciences "Luigi Sacco", Section of Infectious Diseases and Immunopathology, University of Milan, Milano, Italy.
authors.forename: A
authors.initials: A
authors.lastname: Lai
authors.affiliation: Department of Biomedical and Clinical Sciences "Luigi Sacco", Section of Infectious Diseases and Immunopathology, University of Milan, Milano, Italy.
authors.forename: G
authors.initials: G
authors.lastname: Zehender
authors.affiliation: Department of Applied Science and Technology, Politecnico di Torino, Corso Duca degli Abruzzi 24, Torino I-10129, Italy.
authors.forename: D
authors.initials: D
authors.lastname: Mombello
authors.affiliation: Department of Applied Science and Technology, Politecnico di Torino, Corso Duca degli Abruzzi 24, Torino I-10129, Italy; CNR-IMEM, Parco Area delle Scienze 37a, 43124 Parma, Italy.
authors.forename: M
authors.initials: M
authors.lastname: Cocuzza
authors.affiliation: Department of Applied Science and Technology, Politecnico di Torino, Corso Duca degli Abruzzi 24, Torino I-10129, Italy; Center for Space Human Robotics@PoliTo, Istituto Italiano di Tecnologia, C. so Trento 21, 10129 Torino, Italy.
authors.forename: C F
authors.initials: CF
authors.lastname: Pirri
authors.affiliation: Fondazione Bruno Kessler, Laboratory of Biomolecular Sequence and Structure Analysis for Health, Via Sommarive 18, I-38123, Povo, Trento, Italy.
authors.forename: C
authors.initials: C
authors.lastname: Pederzolli
mesh_terms: D000327:Adsorption; D004129:Dimethylpolysiloxanes / Q000737:chemistry; D016174:Hepacivirus / Q000737:chemistry*; D006801:Humans; D056656:Lab-On-A-Chip Devices; D016133:Polymerase Chain Reaction; D012367:RNA, Viral / Q000097:blood* / Q000737:chemistry / Q000302:isolation & purification*; D013499:Surface Properties
chemical_list: D004129:Dimethylpolysiloxanes; D012367:RNA, Viral; C013830:baysilon
keywords: Hepatitis C virus; On-chip RT-PCR; On-chip viral RNA purification; PDMS microdevice