Lateral flow system for nucleic acid detection
First Claim
1. A lateral flow assay device for detecting the presence or absence of at least one single-stranded target nucleic acid in a fluid sample, said device having a first and second end and comprising:
- a sample receiving zone at or near said first end for receiving an aliquot of said sample and comprising a porous material having first and second oligonucleotide probes coupled to first and second binding partners, respectively, wherein said probes specifically hybridize to said target nucleic acid to form a complex having said first and second binding partners, said sample receiving zone being in lateral flow contact with a labeling zone comprising a porous material having at least a first visible moiety reversibly bound thereto and coupled to a first ligand which specifically binds to said first binding partner to form a visible complex, said labeling zone being in lateral flow contact with a capture zone comprising a microporous membrane which contains in a portion thereof a first capture moiety immobilized thereto which specifically binds said second binding partner, said capture zone being in lateral flow contact with an absorbent zone positioned at or near the second end of said device, wherein said visible complex is captured by said capture moiety in said portion of the capture zone.
4 Assignments
0 Petitions
Accused Products
Abstract
The invention provides a complete, one-step, fully functional, ready to use lateral flow assay device for the rapid, accurate detection of a target nucleic acid in a fluid sample, wherein the device contains all reagents necessary for the assay in an anhydrous format. The device comprises a sample receiving zone, a labeling zone, and a capture zone. The sample receiving zone may contain one or more oligonucleotides coupled to binding partners and reversibly bound to the capture zone membrane, the labeling zone comprises a visible moiety coupled to a ligand specific for one of the binding partners and reversibly bound to the labeling zone membrane, and the capture zone comprises an capture moiety specific for the second binding partner and immobilized on the capture zone membrane.
-
Citations
45 Claims
-
1. A lateral flow assay device for detecting the presence or absence of at least one single-stranded target nucleic acid in a fluid sample, said device having a first and second end and comprising:
-
a sample receiving zone at or near said first end for receiving an aliquot of said sample and comprising a porous material having first and second oligonucleotide probes coupled to first and second binding partners, respectively, wherein said probes specifically hybridize to said target nucleic acid to form a complex having said first and second binding partners, said sample receiving zone being in lateral flow contact with a labeling zone comprising a porous material having at least a first visible moiety reversibly bound thereto and coupled to a first ligand which specifically binds to said first binding partner to form a visible complex, said labeling zone being in lateral flow contact with a capture zone comprising a microporous membrane which contains in a portion thereof a first capture moiety immobilized thereto which specifically binds said second binding partner, said capture zone being in lateral flow contact with an absorbent zone positioned at or near the second end of said device, wherein said visible complex is captured by said capture moiety in said portion of the capture zone. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25)
-
-
26. A lateral flow assay device for detection of the presence or absence of at least one target nucleic acid in a fluid sample, wherein said target nucleic acid is coupled to a first binding partner, said device comprising a test strip having a first and second end and comprising:
-
a sample receiving zone at or near said first end for receiving an aliquot of said sample and comprising a porous material having an oligonucleotide probe coupled to a second binding partner, wherein said probe is reversibly bound to said microporous material and specifically hybridizes to said target nucleic acid to form a complex comprising said first and second binding partners, said sample receiving zone being in lateral flow contact with a labeling zone comprising a porous material having at least a first visible moiety reversibly bound thereto and coupled to a first ligand which specifically binds to said first binding partner to form a Visible complex, said labeling zone being in lateral flow contact with a capture zone comprising a microporous membrane which contains in a portion thereof at least a first capture moiety immobilized thereto which specifically binds said second binding partner, said capture zone being in lateral flow contact with an absorbent zone positioned at or near the second end of said test strip, wherein said visible complex is captured by said capture moiety in said portion of the capture zone. - View Dependent Claims (27, 33, 34)
-
-
28. A lateral flow assay device for detection of the presence or absence of at least one target nucleic acid in a fluid sample, wherein said target nucleic acid is coupled to a first and second binding partner, said device comprising a test strip having a first and second end and comprising:
-
a sample receiving zone at or near said first end of said test strip for receiving an aliquot of said sample and comprising a porous material, said sample receiving zone being in lateral flow contact with a labeling zone comprising a porous material having at least a first visible moiety coupled to a first ligand which specifically binds to said first binding partner to form a visible complex, said labeling zone being in lateral flow contact with a capture zone comprising a membrane which contains in at least a portion thereof at least a first capture moiety immobilized thereon which specifically binds said second binding partner, said capture zone being in lateral flow contact with an absorbent zone at or near said second end of said test strip, wherein said visible complex is captured by said capture moiety in said portion of the capture zone. - View Dependent Claims (29)
-
-
30. A method for detecting the presence or absence of at least one target nucleic acid in a fluid sample, said method comprising:
-
(a) applying said sample to a sample receiving zone of a lateral flow test strip of a lateral flow assay device, wherein prior to said application said nucleic acid present in a double-stranded form are rendered single-stranded, wherein said sample wicks sequentially from said sample receiving zone to a labeling zone and to a capture zone of said test strip, said sample receiving zone comprising first and second oligonucleotide probes coupled to first and second binding partners, respectively, and reversibly bound to said test strip, wherein said probes are released from said test strip and specifically hybridize to said target nucleic acid upon contact with said sample to form a complex comprising said first and second binding partners, said labeling zone comprising at least a first visible moiety coupled to a first ligand and reversibly bound to said test strip, wherein said first ligand specifically binds said first binding partner, and said capture zone comprising a capture moiety immobilized on a portion of said test strip, wherein said capture moiety specifically binds said second binding partner; and
(b) detecting the presence of said first visible moiety in said portion of said capture zone. - View Dependent Claims (31, 32, 35, 36, 37, 38, 39)
-
-
40. A method for detecting the presence or absence of at least one target nucleic acid in a fluid sample, said method comprising:
-
(a) coupling said target nucleic acid to a first binding partner to provide a labeled target nucleic acid;
(b) applying said labeled target nucleic acid to a sample receiving zone of a lateral flow test strip of a lateral flow assay device, wherein prior to said application said nucleic acid present in a double-stranded form are rendered single-stranded, wherein said labeled target nucleic acid wicks sequentially from said sample receiving zone to a labeling zone and to a capture zone of said test strip, said sample receiving zone comprising an oligonucleotide probe coupled to a second binding partner and reversibly bound to said test strip, wherein said probes are released from said test strip and specifically hybridize to said labeled target nucleic acid upon contact with said labeled target nucleic acid to form a complex comprising said first and second binding partners, said labeling zone comprising at least a first visible moiety coupled to a first ligand and reversibly bound to said test strip, wherein said first ligand specifically binds said first binding partner, and said capture zone comprising a capture moiety immobilized on a portion of said test strip, wherein said capture moiety specifically binds said second binding partner; and
(c) detecting the presence of said first visible moiety in said portion of said capture zone. - View Dependent Claims (41, 42)
-
-
43. A method for detecting the presence or absence of at least one target nucleic acid in a fluid sample, said method comprising the steps of:
-
(a) coupling said target nucleic acid to a first and second binding partner to provide a labeled target nucleic acid;
(b) applying said labeled target nucleic acid to a sample receiving zone of a lateral flow test strip of a lateral flow assay device, wherein said labeled target nucleic acid laterally wicks sequentially from said sample receiving zone through a labeling zone to a capture zone of said test strip, said labeling zone comprising at least a first visible moiety reversibly bound to said test strip and coupled to a first ligand, wherein said first ligand specifically binds said first binding partner, and said capture zone comprising a capture moiety immobilized on a portion of said test strip, wherein said capture moiety specifically binds said second binding partner; and
(c) detecting the presence of said first visible moiety in said portion of said capture zone. - View Dependent Claims (44, 45)
-
Specification