Point-of-Care System for HTLV-1 Proviral Load Quantification by Digital Mediator Displacement LAMP
Konferenz: MikroSystemTechnik Kongress 2021 - Kongress
08.11.2021 - 10.11.2021 in Stuttgart-Ludwigsburg, Deutschland
Tagungsband: MikroSystemTechnik Kongress 2021
Seiten: 4Sprache: EnglischTyp: PDF
Autoren:
Becherer, Lisa; Hess, Jacob Friedrich; Haerpfer, Tamara; Huber, Dominik; Hutzenlaub, Tobias; Paust, Nils; Borst, Nadine; Zengerle, Roland; von Stetten, Felix (Hahn-Schickard, Freiburg, Germany & Laboratory for MEMS Applications, IMTEK - Department of Microsystems Engineering, University of Freiburg, Freiburg, Germany)
Frischmann, Sieghard; Bakheit, Mohammed (Mast Diagnostica GmbH, Reinfeld, Germany)
Nitschko, Hans; Stinco, Silvina (Department of Virology, Max von Pettenkofer-Institute, Munich, Germany)
Zitz, Friedrich; Hofer, Hannes (E.L.T. Kunststofftechnik & Werkzeugbau GmbH, Friedberg, Austria)
Porro, Giampiero (Datamed srl, Peschiera Borromeo MI, Italy)
Hausladen, Florian; Stock, Karl; Drossart, Dominik; Wurm, Holger (Institut für Lasertechnologien in der Medizin und Meßtechnik, University of Ulm, Ulm, Germany)
Kuhn, Hanna (Hahn-Schickard, Freiburg, Germany & NB Technologies GmbH, Bonn, Germany)
Keller, Mark; Strohmeier, Oliver (Hahn-Schickard, Freiburg, Germany & Laboratory for MEMS Applications, IMTEK - Department of Microsystems Engineering, University of Freiburg, Freiburg, Germany & Spindiag GmbH, Freiburg, Germany)
Wadle, Simon (Hahn-Schickard, Freiburg, Germany & Laboratory for MEMS Applications, IMTEK - Department of Microsystems Engineering, University of Freiburg, Freiburg, Germany & Volpi AG, Schlieren, Switzerland)
Inhalt:
We present a universal point-of-care (POC) system for fully automated quantification of human T-cell lymphotropic virus type 1 (HTLV-1) proviral load, including genomic RNA, based on digital mediator displacement LAMP (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 (run time of two hours and 10 min).