Fiber optic biosensor for selectively detecting oligonucleotide species in a mixed fluid sample

• US 6,482,593 B2
• Filed: 11/05/1998
• Issued: 11/19/2002
• Est. Priority Date: 05/05/1997
• Status: Expired due to Term

• Current Assignee: Trustees of Tufts College
• Original Assignee: Trustees of Tufts College
• Inventors: Healey, Brian G., Walt, David R.
• Primary Examiner(s): FREDMAN, JEFFREY NORMAN
• Application Number:
  US09/187,289
• Publication Number:
  US 20020009719A1
• Time in Patent Office: 1,475 Days
• Field of Search: 435/6, 435/91.2, 536/23.1, 536/24.3, 422/55, 422/68.1, 422/82.05, 422/82.06, 422/82.07, 422/82.08, 654/09
• US Class Current: 506/17
• CPC Class Codes: G01N 21/7703 using reagent-clad optical fibres or optical waveguides using measurement of total internal reflection or attenuated total reflection G01N21/552; optical fibres or waveguides per se G02B
• CPC Class Codes: G01N 2201/0833 Fibre array at detector, resolving
• CPC Class Codes: Y10S 977/924 using nanostructure as support of dna analysis

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 species of single stranded nucleic acid disposed as individual deposits in aligned organization upon multiple strand end faces at differing spatial positions on the distal array end surface.

Claims

1. An optical sensor for detecting nucleic acids in a fluid sample comprising:

a) 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 distal end, each of said ends being formed by multiple strand faces of said strands, at least one of said array ends presenting a discrete fiber optic array surface for introduction and conveyance of light energy; and

b) at least a first and a second nucleic acid attached to a first and second portion, respectively, of said distal array end, said first nucleic acid being optically coupled to and in optical communication with a first of said multiple end faces at said distal array end, and said second nucleic acid being optically coupled to and in optical communication with a second of said multiple end faces at said distal array end.

2. A sensor according to claim 1 wherein said nucleic acid is selected from the group consisting of DNA, RNA and PNA.

3. A sensor according to claim 1 wherein said preformed unitary 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.

4. A sensor according to claim 1 further comprising a light source.

5. A sensor according to claim 1 further comprising a light detector.

6. A sensor according to claim 1 wherein said nucleic acid is attached to said portion by introducing light energy to at least said first end face in the presence of a photoactivatable compound.

7. A sensor according to claim 6 wherein said photoactivatable compound is selected from the group consisting of acrylamide, N,N-methylene bis(acrylamide), hydroxyehtylmethacrylate, EGDMA, vinyl acetate, N-(3-aminopropyl)methacrylamide, hydrochloride, and N-acryloxysuccinimide.

8. A sensor according to claim 6 wherein said light energy is introduced at said proximal end.

9. A sensor according to claim 6 wherein said light energy is introduced at said distal end.

10. A method of making a fiber optic sensor comprising:

a) providing a preformed, unitary fiber optic array comprising a plurality of optic fiber strands disposed co-axially and joined along their lengths, said fiber optic array having a proximal and a distal array end, each of said ends being formed of multiple strand end faces and at least one of said array ends presenting a discrete optic array surface for introduction and conveyance of light energy;

b) contacting at least a first nucleic acid and a photoactivatable compound with at least a first proximal optic fiber strand end face; and

c) attaching said first nucleic acid to said end face by introducing light energy to said first optic fiber strand.

11. A method according to claim 10 further comprising:

d) contacting at least a second nucleic acid and a photoactivatable compound with at least a second proximal first optic fiber strand end face; and

e) attaching said second nucleic acid to said second end face by introducing light energy to said second optic fiber strand.

12. A method according to claim 10 wherein said light energy is introduced into the first optic fiber strand at the distal end.

13. A method according to claim 10 wherein said light energy is introduced at said proximal end.

14. A method according to claim 10 wherein said nucleic acid is selected from the group consisting of DNA, RNA and PNA.

15. A method according to claim 10 wherein said introduction utilizes a light source and a mask for illuminating a portion of said array end surface for precisely controlling the location and size of said nucleic acid attachment.

16. A method of detecting at least one target sequence in a fluid sample comprising:

a) providing a a preformed, unitary fiber optic array comprising a plurality of optic fiber strands disposed co-axially and joined along their lengths, said fiber optic array having a proximal and a distal array end, each of said ends being formed of multiple strand end faces and at least one of said array ends presenting a discrete optic array surface for introduction and conveyance of light energy, said distal array end comprising a plurality of nucleic acid probes, each of which is attached to a different strand end face;

b) contacting said distal end of said array with said sample; and

c) detecting the presence of a target sequence.

17. A method according to claim 16 wherein said target sequence is labeled and said detecting is done by illuminating said label with excitation light energy and detecting emission light energy from said label.

18. A method according to claim 17 wherein said illuminating is done at said proximal end of said array.

19. A method according to claim 17 wherein said illuminating is done at said distal end of said array.