Miniaturized lateral flow device for rapid and sensitive detection of proteins or nucleic acids
First Claim
1. A device for detecting the presence of a target nucleic acid in a fluid sample, the device comprising:
- a porous lateral flow strip having a width of 3 mm or less; and
a microarray of spots on said porous lateral flow strip, each spot comprising a plurality of capture oligonucleotides;
wherein each capture oligonucleotide is complementary to a capture sequence of the target nucleic acid;
wherein each spot was printed with a solution having a concentration of capture oligonucleotides between 200 μ
M and 800 μ
M; and
wherein said microarray of spots provides a quantitative visual colorimetric signal indicating an amount of from 250 amol to 1 fmol of the target nucleic acid in the fluid sample comprising at least 150 mM salt in less than two minutes by capillary flow.
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Abstract
The invention provides miniaturized lateral flow chromatographic and lateral flow chromatographic microarray devices (LFM). The miniaturization of lateral flow nucleic acid detection achieved by the present invention offers reduced reagent use, femtomole sensitivity, excellent linear dynamic range, and rapid detection. Moreover, the small feature sizes of capture oligonucleotides renders the potential information capacity of the platform comparable to more traditional spotted fluorescence microarrays as well as improving sensitivity. The LFM devices exemplified herein enable analytes to be detected within 10 seconds from the time of sample introduction to the LFM device. Sample volumes may be as low as about 10 microliters, significantly reducing assay costs and ameliorating reagent storage logistics. Additionally, the miniaturization of lateral flow opens the door to highly multiplexed assays, allowing many proteins or nucleic acids to be detected in a single assay.
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Citations
18 Claims
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1. A device for detecting the presence of a target nucleic acid in a fluid sample, the device comprising:
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a porous lateral flow strip having a width of 3 mm or less; and a microarray of spots on said porous lateral flow strip, each spot comprising a plurality of capture oligonucleotides; wherein each capture oligonucleotide is complementary to a capture sequence of the target nucleic acid; wherein each spot was printed with a solution having a concentration of capture oligonucleotides between 200 μ
M and 800 μ
M; andwherein said microarray of spots provides a quantitative visual colorimetric signal indicating an amount of from 250 amol to 1 fmol of the target nucleic acid in the fluid sample comprising at least 150 mM salt in less than two minutes by capillary flow. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18)
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Specification