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

Becherer, Lisa; Hess, Jacob Friedrich; Frischmann, Sieghard; Bakheit, Mohammed; Nitschko, Hans; Stinco, Silvina; Zitz, Friedrich; Hofer, Hannes; Porro, Giampiero; Hausladen, Florian; Stock, Karl; Drossart, Dominik; Wurm, Holger; Kuhn, Hanna; Huber, Dominik; Hutzenlaub, Tobias; Paust, Nils; Keller, Mark; Strohmeier, Oliver; Wadle, Simon; Borst, Nadine; Zengerle, Roland and Stetten, Felix von (2021): Point-of-Care System for HTLV-1 Proviral Load Quantification by Digital Mediator Displacement LAMP. In: Micromachines, Vol. 12, No. 2, 159

Abstract

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.

• Item Type: Journal article
• Faculties: Medicine
• Subjects: 600 Technology > 610 Medicine and health
• Language: English