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Department of Electrical Engineering, University of Washington, Seattle, USA
E-mail: llafleur@u.washington.edu
; Fax: +01 (206)543-3842
; Tel: +01 (206)616-1928
b
Department of Bioengineering, University of Washington, Seattle, USA
c
Department of Chemistry, University of Washington, Seattle, USA
d
PATH, Seattle, USA
e
Oxford University Clinical Research Unit-Patan Academy of Health Sciences, Kathmandu, Nepal
Lab Chip, 2012,12, 1119-1127
DOI:
10.1039/C2LC20751F
Received
12 Aug 2011,
Accepted
10 Jan 2012
First published online
07 Feb 2012
In many low resource settings multiple diseases are endemic. There is a need for appropriate multi-analyte diagnostics capable of differentiating between diseases that cause similar clinical symptoms. The work presented here was part of a larger effort to develop a microfluidic point-of-care system, the DxBox, for sample-to-result differential diagnosis of infections that present with high rapid-onset fever. Here we describe a platform that detects disease-specific antigens and IgM antibodies. The disposable microfluidic cards are based on a flow-through membrane immunoassay carried out on porous nitrocellulose, which provides rapid diffusion for short assay times and a high surface area for visual detection of colored assay spots. Fluid motion and on-card valves were driven by a pneumatic system and we present designs for using pneumatic control to carry out assay functions. Pneumatic actuation, while having the potential advantage of inexpensive and robust hardware, introduced bubbles that interfered with fluidic control and affected assay results. The cards performed all sample preparation steps including plasma filtration from whole blood, sample and reagent aliquoting for the two parallel assays, sample dilution, and IgG removal for the IgM assays. We demonstrated the system for detection of the malarial pfHRPII antigen (spiked) and IgM antibodies to Salmonella Typhi LPS (patient plasma samples). All reagents were stored on card in dry form; only the sample and buffer were required to run the tests. Here we detail the development of this platform and discuss its strengths and weaknesses.
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