Point-of-Care System for HTLV-1 Proviral Load Quantification by Digital Mediator Displacement LAMP.

Autor:	Becherer L; Hahn-Schickard, Georges-Koehler-Allee 103, 79110 Freiburg, Germany.; Laboratory for MEMS Applications, IMTEK-Department of Microsystems Engineering, University of Freiburg, Georges-Koehler-Allee 103, 79110 Freiburg, Germany., Hess JF; Hahn-Schickard, Georges-Koehler-Allee 103, 79110 Freiburg, Germany.; Laboratory for MEMS Applications, IMTEK-Department of Microsystems Engineering, University of Freiburg, Georges-Koehler-Allee 103, 79110 Freiburg, Germany., Frischmann S; Mast Diagnostica GmbH, Feldstraße 20, 23858 Reinfeld, Germany., Bakheit M; Mast Diagnostica GmbH, Feldstraße 20, 23858 Reinfeld, Germany., Nitschko H; Department of Virology, Pettenkoferstraße 9a, Max von Pettenkofer-Institute, 80336 Munich, Germany., Stinco S; Department of Virology, Pettenkoferstraße 9a, Max von Pettenkofer-Institute, 80336 Munich, Germany., Zitz F; E.L.T. Kunststofftechnik & Werkzeugbau GmbH, Weidenweg 339, 8240 Friedberg, Austria., Hofer H; E.L.T. Kunststofftechnik & Werkzeugbau GmbH, Weidenweg 339, 8240 Friedberg, Austria., Porro G; Datamed srl, Via Achille Grandi 4/6, 20068 Peschiera Borromeo MI, Italy., Hausladen F; Institut für Lasertechnologien in der Medizin und Meßtechnik, University of Ulm, Helmholtzstraße 12, 89081 Ulm, Germany., Stock K; Institut für Lasertechnologien in der Medizin und Meßtechnik, University of Ulm, Helmholtzstraße 12, 89081 Ulm, Germany., Drossart D; Institut für Lasertechnologien in der Medizin und Meßtechnik, University of Ulm, Helmholtzstraße 12, 89081 Ulm, Germany., Wurm H; Institut für Lasertechnologien in der Medizin und Meßtechnik, University of Ulm, Helmholtzstraße 12, 89081 Ulm, Germany., Kuhn H; Hahn-Schickard, Georges-Koehler-Allee 103, 79110 Freiburg, Germany.; NB Technologies GmbH, Ludwig-Erhard-Allee 2, 53175 Bonn, Germany., Huber D; Hahn-Schickard, Georges-Koehler-Allee 103, 79110 Freiburg, Germany.; Laboratory for MEMS Applications, IMTEK-Department of Microsystems Engineering, University of Freiburg, Georges-Koehler-Allee 103, 79110 Freiburg, Germany., Hutzenlaub T; Hahn-Schickard, Georges-Koehler-Allee 103, 79110 Freiburg, Germany.; Laboratory for MEMS Applications, IMTEK-Department of Microsystems Engineering, University of Freiburg, Georges-Koehler-Allee 103, 79110 Freiburg, Germany., Paust N; Hahn-Schickard, Georges-Koehler-Allee 103, 79110 Freiburg, Germany.; Laboratory for MEMS Applications, IMTEK-Department of Microsystems Engineering, University of Freiburg, Georges-Koehler-Allee 103, 79110 Freiburg, Germany., Keller M; Hahn-Schickard, Georges-Koehler-Allee 103, 79110 Freiburg, Germany.; Laboratory for MEMS Applications, IMTEK-Department of Microsystems Engineering, University of Freiburg, Georges-Koehler-Allee 103, 79110 Freiburg, Germany.; Spindiag GmbH, Engesserstraße 4a, 79108 Freiburg, Germany., Strohmeier O; Hahn-Schickard, Georges-Koehler-Allee 103, 79110 Freiburg, Germany.; Laboratory for MEMS Applications, IMTEK-Department of Microsystems Engineering, University of Freiburg, Georges-Koehler-Allee 103, 79110 Freiburg, Germany.; Spindiag GmbH, Engesserstraße 4a, 79108 Freiburg, Germany., Wadle S; Hahn-Schickard, Georges-Koehler-Allee 103, 79110 Freiburg, Germany.; Laboratory for MEMS Applications, IMTEK-Department of Microsystems Engineering, University of Freiburg, Georges-Koehler-Allee 103, 79110 Freiburg, Germany.; Volpi AG, Wiesenstrasse 33, 8952 Schlieren, Switzerland., Borst N; Hahn-Schickard, Georges-Koehler-Allee 103, 79110 Freiburg, Germany.; Laboratory for MEMS Applications, IMTEK-Department of Microsystems Engineering, University of Freiburg, Georges-Koehler-Allee 103, 79110 Freiburg, Germany., Zengerle R; Hahn-Schickard, Georges-Koehler-Allee 103, 79110 Freiburg, Germany.; Laboratory for MEMS Applications, IMTEK-Department of Microsystems Engineering, University of Freiburg, Georges-Koehler-Allee 103, 79110 Freiburg, Germany., von Stetten F; Hahn-Schickard, Georges-Koehler-Allee 103, 79110 Freiburg, Germany.; Laboratory for MEMS Applications, IMTEK-Department of Microsystems Engineering, University of Freiburg, Georges-Koehler-Allee 103, 79110 Freiburg, Germany.
Jazyk:	angličtina
Zdroj:	Micromachines [Micromachines (Basel)] 2021 Feb 05; Vol. 12 (2). Date of Electronic Publication: 2021 Feb 05.
DOI:	10.3390/mi12020159
Abstrakt:	This paper presents a universal point-of-care system for fully automated quantification of human T-cell lymphotropic virus type 1 (HTLV-1) proviral load, including genomic RNA, based on digital reverse RNA transcription and c-DNA amplification by MD LAMP (mediator displacement loop-mediated isothermal amplification). A disposable microfluidic LabDisk with pre-stored reagents performs automated nucleic acid extraction, reaction setup, emulsification, reverse transcription, digital DNA amplification, and quantitative fluorogenic endpoint detection with universal reporter molecules. Automated nucleic acid extraction from a suspension of HTLV-1-infected CD4+ T-lymphocytes (MT-2 cells) yielded 8 ± 7 viral nucleic acid copies per MT-2 cell, very similar to the manual reference extraction (7 ± 2 nucleic acid copies). Fully automated sample processing from whole blood spiked with MT-2 cells showed a comparable result of 7 ± 3 copies per MT-2 cell after a run time of two hours and 10 min.
Databáze:	MEDLINE
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