Point-of-Care System for HTLV-1 Proviral Load Quantification by Digital Mediator Displacement LAMP.
centrifugal microfluidics
digital loop-mediated isothermal amplification
human T-cell lymphotropic virus 1
point-of-care testing
proviral load measurements
Journal
Micromachines
ISSN: 2072-666X
Titre abrégé: Micromachines (Basel)
Pays: Switzerland
ID NLM: 101640903
Informations de publication
Date de publication:
05 Feb 2021
05 Feb 2021
Historique:
received:
18
01
2021
revised:
29
01
2021
accepted:
02
02
2021
entrez:
10
2
2021
pubmed:
11
2
2021
medline:
11
2
2021
Statut:
epublish
Résumé
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.
Identifiants
pubmed: 33562822
pii: mi12020159
doi: 10.3390/mi12020159
pmc: PMC7915047
pii:
doi:
Types de publication
Journal Article
Langues
eng
Subventions
Organisme : Bundesministerium für Bildung und Forschung
ID : 031B0132B
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