Abstract
Preliminary development of a simple mesofluidic multi-channel plastic cartridge with underlying external magnet to drag DNA aptamer-coated paramagnetic beads through fluids in the channels while developing a sandwich assay with quantum dot-conjugated reporter aptamers is described. This approach is superior to traditional lateral flow test strips in several ways including: 1) the ability to control the speed of lateral flow in the channel versus conventional nitrocellulose analytical membranes with fixed wicking times. 2) The use of aptamers for potentially greater affinity and consistency from batch-to-batch versus comparable antibodies. 3) Superior fluorescence efficiency and intensity provided by quantum dots versus conventional fluorescent dyes and 4) the ability to multiplex based on the various colored emissions of different sized quantum dots when excited with a single ultraviolet source. Development of the system from concept to prototype is described along with illustration of sensitive system performance for several food safety-related targets. The system is also clearly adaptable to rapid multiplex detection and sensitive quantitation of clinical biomarkers, drugs, environmental, veterinary or other target analytes.
Highlights
Lateral flow (LF) or immunochromatographic (IC) test strips have long dominated the rapid detection markets in the areas of food safety, clinical, drug and other testing for their convenience, speed, portability, minimal training requirements, and low cost which is especially important in resource-limited areas or underdeveloped countries [1]
Our group has demonstrated that aptamers it has developed against Campylobacter jejuni and Listeria monocytogenes can be used in quantum dots (QDs) or enzymelinked fluorescence aptamer-paramagnetic bead (MB) sandwich assays to achieve detection limits below 10 colony forming units or cells per ml [13,15]
We describe our first attempts to develop a multiplexed cartridge in which various aptamerMB plus aptamer-QD sandwich assays are added to multiple linear mesofluidic channels in a plastic cartridge or tray for simultaneous multiplex testing
Summary
Lateral flow (LF) or immunochromatographic (IC) test strips have long dominated the rapid detection markets in the areas of food safety, clinical (especially pregnancy), drug and other testing for their convenience, speed, portability, minimal training requirements, and low cost which is especially important in resource-limited areas or underdeveloped countries [1]. We describe our first attempts to develop a multiplexed cartridge in which various aptamerMB plus aptamer-QD sandwich assays are added to multiple linear mesofluidic channels in a plastic cartridge or tray for simultaneous multiplex testing. These assays are assisted by an external magnet to traverse the channels while the sandwich assays develop and are separated from and washed free of excess aptamer-QD conjugates and any potential food particles or other debris to generate a purified fluorescent assay on MBs for visual or fluorometric detection with a single UV excitation source [33]
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