Miniaturized lateral flow device for rapid and sensitive detection of proteins or nucleic acids
First Claim
1. A lateral flow chromatographic device for detecting the presence of at least one single-stranded target nucleic acid analyte in a fluid sample, comprising a chromatographic test strip which comprises (a) a sample receiving zone for receiving an aliquot of the sample and for receiving a labeled detection oligonucleotide, which detection oligonucleotide comprises a sequence which is complementary to a first sequence of the target nucleic acid;
- and, (b) a capture zone in lateral flow contact with the sample receiving zone, said capture zone comprising a microporous membrane, onto which at least one capture oligonucleotide is immobilized at a feature size of 500 μ
m diameter or smaller, and which comprises a sequence which is complementary to a second sequence of the target nucleic acid.
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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
42 Claims
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1. A lateral flow chromatographic device for detecting the presence of at least one single-stranded target nucleic acid analyte in a fluid sample, comprising a chromatographic test strip which comprises (a) a sample receiving zone for receiving an aliquot of the sample and for receiving a labeled detection oligonucleotide, which detection oligonucleotide comprises a sequence which is complementary to a first sequence of the target nucleic acid;
- and, (b) a capture zone in lateral flow contact with the sample receiving zone, said capture zone comprising a microporous membrane, onto which at least one capture oligonucleotide is immobilized at a feature size of 500 μ
m diameter or smaller, and which comprises a sequence which is complementary to a second sequence of the target nucleic acid. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 29, 30, 31)
- and, (b) a capture zone in lateral flow contact with the sample receiving zone, said capture zone comprising a microporous membrane, onto which at least one capture oligonucleotide is immobilized at a feature size of 500 μ
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15. A lateral flow chromatographic device for detecting the presence of at least one single-stranded target nucleic acid analyte in a fluid sample, comprising a lateral flow matrix which defines a flow path and which comprises in series:
- (a) a sample receiving zone for receiving an aliquot of a fluid sample;
(b) a labeling zone in lateral flow contact with said sample receiving zone, wherein the labeling zone comprises a porous material containing at least one detection oligonucleotide reversibly bound thereto, which detection oligonucleotide is complementary to a first sequence of the target nucleic acid and is coupled to a detectable label; and
, (c) a capture zone in lateral flow contact with said labeling zone, said capture zone comprising a microporous membrane, onto which at least one capture oligonucleotide is immobilized at a feature size of 500 μ
m diameter or smaller. - View Dependent Claims (16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 32, 33, 34)
- (a) a sample receiving zone for receiving an aliquot of a fluid sample;
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35. A lateral flow microarray chromatographic device for detecting the presence or absence of a plurality of single-stranded target nucleic acids in one or more fluid samples, comprising a lateral flow matrix which defines a flow path and which comprises in series:
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(a) a sample receiving zone for receiving the fluid sample(s); (b) a labeling zone in lateral flow contact with said sample receiving zone, wherein the labeling zone comprises a porous material containing a plurality of different detection oligonucleotides reversibly bound thereto, which detection oligonucleotides are complementary to first sequences of a plurality of respective target nucleic acids and are coupled to detectable labels; and
,(c) a capture zone in lateral flow contact with said labeling zone, said capture zone comprising a microporous membrane, at least a portion of which contains a plurality of different capture oligonucleotides immobilized thereto, which capture oligonucleotides are complementary to second sequences of a plurality of respective target nucleic acids, and wherein the different capture oligonucleotides are immobilized to the microporous membrane at a feature size of 300 μ
m or less in diameter. - View Dependent Claims (36, 37, 38)
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39. A lateral flow microarray chromatographic device for detecting the presence or absence of a plurality of target nucleic acids in one or more fluid samples, comprising a lateral flow matrix which defines a flow path and which comprises in series:
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(a) a sample receiving zone for receiving the fluid sample(s) and for receiving a plurality of different detection oligonucleotides, each of which detection oligonucleotides comprises a sequence which is complementary to a first sequence of a specific target nucleic acid and is labeled; and
,(b) a capture zone in lateral flow contact with said labeling zone, said capture zone comprising a microporous membrane, at least a portion of which contains a plurality of different capture oligonucleotides immobilized thereto, each of which capture oligonucleotides comprises a sequence which is complementary to second sequence of the specific target nucleic acid, and wherein the different capture oligonucleotides are immobilized to the microporous membrane at a feature size of 300 μ
m or less in diameter. - View Dependent Claims (40, 41, 42)
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Specification