LabDisk with complete reagent prestorage for sample-to-answer nucleic acid based detection of respiratory pathogens verified with Influenza A H3N2 virus

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Abstract: Portable point-of-care devices for pathogen detection require easy, minimal and user-friendly handling steps and need to have the same diagnostic performance compared to centralized laboratories. In this work we present a fully automated sample-to-answer detection of influenza A H3N2 virus in a centrifugal LabDisk with complete prestorage of reagents. Thus, the initial supply of the sample remains the only manual handling step. The self-contained LabDisk automates by centrifugal microfluidics all necessary process chains for PCR-based pathogen detection: pathogen lysis, magnetic bead based nucleic acid extraction, aliquoting of the eluate into 8 reaction cavities, and real-time reverse transcription polymerase chain reaction (RT-PCR). Prestored reagents comprise air dried specific primers and fluorescence probes, lyophilized RT-PCR mastermix and stick-packaged liquid reagents for nucleic acid extraction. Employing two different release frequencies for the stick-packaged liquid reagents enables on-demand release of highly wetting extraction buffers, such as sequential release of lysis and binding buffer. Microfluidic process-flow was successful in 54 out of 55 tested LabDisks. We demonstrate successful detection of the respiratory pathogen influenza A H3N2 virus in a total of 18 LabDisks with sample concentrations down to 2.39 × 104 viral RNA copies per ml, which is in the range of clinical relevance. Furthermore, we detected RNA bacteriophage MS2 acting as internal control in 3 LabDisks with a sample concentration down to 75 plaque forming units (pfu) per ml. All experiments were applied in a 2 kg portable, laptop controlled point-of-care device. The turnaround time of the complete analysis from sample-to-answer was less than 3.5 hours

Location
Deutsche Nationalbibliothek Frankfurt am Main
Extent
Online-Ressource
Language
Englisch
Notes
Lab on a chip 16 (2016), 199-207, doi 10.1039/C5LC00871A, issn: 1473-0189
IN COPYRIGHT http://rightsstatements.org/page/InC/1.0 rs

Classification
Industrielle und handwerkliche Fertigung

Event
Veröffentlichung
(where)
Freiburg
(who)
Albert-Ludwigs-Universität Freiburg
(when)
2016
Creator
Contributor
BIOSS Centre for Biological Signalling Studies
Institut für Mikro- und Informationstechnik der Hahn-Schickard Gesellschaft für angewandte Forschung e.V
Professur für Anwendungsentwicklung
Technische Fakultät
Albert-Ludwigs-Universität Freiburg

DOI
10.1039/C5LC00871A
URN
urn:nbn:de:bsz:25-freidok-121293
Rights
Der Zugriff auf das Objekt ist unbeschränkt möglich.
Last update
14.08.2025, 11:02 AM CEST

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Associated

Time of origin

  • 2016

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