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

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.

Diagnóstico molecular de vírus T-linfotrópico humano (HTLV): histórico e estado da arte

“…Esse fenômeno somente é possível por meio do uso de uma Diagnóstico molecular de vírus T-linfotrópico humano (HTLV): histórico e estado da arte/Caterino-de-Araujo A e Gonçalves MG para quantificação de CPV de HTLV-1. 106 Este último sistema foi idealizado por pesquisadores da Alemanha como uma plataforma de trabalho que se assemelha a uma centrífuga; nele é feita a extração de RNA genômico, transcrição para c-DNA, amplificação e detecção do material amplificado usando a técnica LAMP, com finalização em 2h e 10min 106 . Apesar dessas inovações tecnológicas, elas ainda não se aplicam à realidade brasileira.…”

Programmable magnetic robot (ProMagBot) for automated nucleic acid extraction at the point of need

“…Extraction devices that process PMB kits have shown integrated permanent magnets, electromagnet (EM) controls, EM-actuated magnets, centrifugestyle rotations, automated pipetting, and fluid flow methods. 14,[45][46][47][48][49][50][51][52][53][54][55][56] These methods aim to simplify the handling process for the user. However, these devices lack sample-toextraction automation and still require intermittent manual steps.…”