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2016 ; 1
(4
): 586-599
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Multiplex Flow Assays
#MMPMID27819063
Haushalter KJ
; Vetcha S
; Haushalter RC
ACS Omega
2016[Oct]; 1
(4
): 586-599
PMID27819063
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Lateral flow or dipstick assays (e.g., home pregnancy tests), where an analyte
solution is drawn through a porous membrane and is detected by localization onto
a capture probe residing at a specific site on the flow strip, are the most
commonly and extensively used type of diagnostic assay. However, after over 30
years of use, these assays are constrained to measuring one or a few analytes at
a time. Here, we describe a completely general method, in which any single-plex
lateral flow assay is transformed into a multiplex assay capable of measuring an
arbitrarily large number of analytes simultaneously. Instead of identifying the
analyte by its localization onto a specific geometric location in the flow
medium, the analyte-specific capture probe is identified by its association with
a specific optically encoded region within the flow medium. The capture probes
for nucleic acids, antigens, or antibodies are attached to highly porous agarose
beads, which have been encoded using multiple lanthanide emitters to create a
unique optical signature for each capture probe. The optically encoded capture
probe-derivatized beads are placed in contact with the analyte-containing porous
flow medium and the analytes are captured onto the encoded regions as the
solution flows through the porous medium. To perform a multiplex diagnostic
assay, a solution comprising multiple analytes is passed through the flow medium
containing the capture probe-derivatized beads, and the captured analyte is
treated with a suitable fluorescent reporter. We demonstrate this multiplex
analysis technique by simultaneously measuring DNA samples, antigen-antibody
pairs, and mixtures of multiple nucleic acids and antibodies.
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