Fiber optic biosensor for selectively detecting oligonucleotide species in a mixed fluid sample
First Claim
1. An optical sensor for detecting nucleic aids in a fluid sample comprising:
- a preformed, unitary fiber optic array comprising a plurality of optical fiber strands, said strands being disposed co-axially and joined along their lengths, said fiber optic array having a proximal and a distal end;
a polymer immobilized to at least a first and second portion of said distal end; and
at least a first and a second nucleic acid attached to said polymer immobilized to at least a first and second portion of said distal end.
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Accused Products
Abstract
The present invention provides biosensors, apparatus and methods for selectively detecting at least one complementary oligonucleotide target specie in a fluid sample containing a mixture of different oligonucleotide fragments. One preferred embodiment of the biosensor is as a unitary fiber optic array having an in-situ hybridization zone comprising not less than one specie of single stranded oligonucleotide disposed as individual deposits in aligned organization upon multiple strand end faces at differing spatial positions on the distal array end surface. In this manner, a collective of deployed, single specie, multiple fixed probes are presented for selective in-situ hybridization on-demand with at least one mobile complementary target specie ultimately bearing a joined identifying label. The biosensor provides for optical detection of in-situ hybridization on the distal end surface via the presence of the concomitantly disposed joined identifying label at the differing spatial positions.
167 Citations
30 Claims
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1. An optical sensor for detecting nucleic aids in a fluid sample comprising:
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a preformed, unitary fiber optic array comprising a plurality of optical fiber strands, said strands being disposed co-axially and joined along their lengths, said fiber optic array having a proximal and a distal end;
a polymer immobilized to at least a first and second portion of said distal end; and
at least a first and a second nucleic acid attached to said polymer immobilized to at least a first and second portion of said distal end. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18)
at least one second nucleic acid attached to a polymer immobilized to at least one second portion of said distal array end.
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11. A sensor according to claim 10 wherein said nucleic acid is selected from the group consisting of DNA, RNA and PNA.
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12. A sensor according to claim 10 wherein said preformed, unitary fiber optic array is selected from the group consisting of an imaging fiber, a coherent fiber array, a random fiber array, and a semi-random fiber array.
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13. A sensor according to claim 10 further comprising a light source.
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14. The sensor according to claim 13 further comprising a light detector.
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15. A sensor according to claim 10 wherein said first nucleic acid is attached to said polymer immobilized to said first portion of said distal end by introducing light energy to said multiple strand end faces at a first portion of said proximal array end, said light energy introduced in the presence of a photactivatable compound.
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16. A sensor according to claim 15 wherein said photoactivatable compound is selected from the group consisting of acrylamide, N,N-methylene bis (acrylamide), hydroxyethylmethacrylate, EGDMA, vinyl acetate, N-(3-aminopropyl) methacrylamide, hydrochloride, and N-acryloxy succinimide.
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17. A sensor according to claim 10 wherein said first nucleic acid is attached to said polymer immobilized to said first portion of said distal end by introducing light energy to said multiple strand end faces at a first portion of said distal array end, said light energy introduced in the presence of a photactivatable compound.
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18. A sensor according to claim 17 wherein said photoactivatable compound is selected from the group consisting of acrylamide, N,N-methylene bis (acrylamide), hydroxyethylmethacrylate, EGDMA, vinyl acetate, N-(3-aminopropyl) methacrylamide, hydrochloride, and N-acryloxy succinimide.
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19. A method of detecting at least one nucleic acid in a fluid sample comprising the steps of:
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a) providing a preformed, unitary fiber optic array comprising;
i) a plurality of optical fiber strands, said strands being disposed co-axially and joined along their lengths, said fiber optic array having a proximal and a distal end; and
ii) at least one first nucleic acid attached to a first polymer immobilized to a portion of said distal array end;
b) contacting said distal end of said array with said sample fluid comprising at least one labeled complementary target nucleic acid; and
c) detecting a complex comprised of said at least one nucleic acid and said at least one labeled target nucleic acid. - View Dependent Claims (20, 21, 22)
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23. A method of making a fiber optic sensor comprising:
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a) providing a unitary fiber optic array comprising a plurality of optical fiber strands b) immobilizing a polymer to at least a first distal optical fiber strand end face by introducing light energy to said first optical fiber strand end face; and
c) attaching said first nucleic acid to said immobilized polymer. - View Dependent Claims (24, 25, 26, 27, 28, 29, 30)
d) immobilizing a polymer to at least a second distal optical fiber strand end face by introducing light energy to said optical fiber strand end face; and
e) attaching said second nucleic acid to said second immobilized polymer.
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29. A method according to claim 28 wherein said nucleic acid is selected from the group consisting of DNA, RNA and PNA.
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30. A method according to claim 28 wherein said introducing light energy further compises employing a light source and a masking means for illuminating a portion of said array end surface for precisely controlling the location and size of said polymer immobilization.
Specification