Surface enhanced Raman gene probe and methods thereof

• US 5,721,102 A
• Filed: 01/31/1997
• Issued: 02/24/1998
• Est. Priority Date: 10/13/1995
• Status: Expired due to Fees

• Current Assignee: Lockheed Martin Energy Systems, Inc. (AES Corporation)
• Original Assignee: Lockheed Martin Energy Systems, Inc. (AES Corporation)
• Inventors: Vo-Dinh, Tuan
• Primary Examiner(s): Campbell, Eggerton A.
• Application Number:
  US08/792,085
• Time in Patent Office: 389 Days
• Field of Search: 435/5, 435/6, 935/77, 935/78
• US Class Current: 435/6.12
• CPC Class Codes: C12Q 1/6825 Nucleic acid detection involving sensors
• CPC Class Codes: C12Q 2565/518 characterised by the immobilisation of the nucleic acid sample or target
• CPC Class Codes: C12Q 2565/632 being a surface enhanced, e.g. resonance, Raman spectrometer
• CPC Class Codes: G01N 21/658 enhancement Raman, e.g. surface plasmons

Abstract

The subject invention disclosed herein is a new gene probe biosensor and methods thereof based on surface enhanced Raman scattering (SERS) label detection. The SER gene probe biosensor comprises a support means, a SER gene probe having at least one oligonucleotide strand labeled with at least one SERS label, and a SERS active substrate disposed on the support means and having at least one of the SER gene probes adsorbed thereon. Biotargets such as bacterial and viral DNA, RNA and PNA are detected using a SER gene probe via hybridization to oligonucleotide strands complementary to the SER gene probe. The support means supporting the SERS active substrate includes a fiberoptic probe, an array of fiberoptic probes for performance of multiple assays and a waveguide microsensor array with charge-coupled devices or photodiode arrays.

Claims

1. A method for using a SER Gene Probe for hybridization, detection and identification of hybridized target oligonucleotide strands comprising the steps of:

a) preparing a sampling medium with immobilized oligonucleotide strands of known sequence adsorbed thereon, said immobilized oligonucleotide strands being complementary to said target oligonucleotide strands;

b) synthesizing SER gene probes wherein a SER gene probe comprises at least one oligonucleotide strand of known sequence having at least one SERS label;

c) preparing a SER gene probe solution comprising at least one SER gene probe wherein said SERS label is unique to said oligonucleotide strand of a particular sequence;

d) incubating said sample medium in an amount of SER gene probe solution sufficient enough to hybridize said immobilized oligonucleotide strands on said sample medium with said target oligonucleotide strands that are complementary to said immobilized oligonucleotide strands, incubating for a time period sufficient enough as for said SER gene probes to contact said immobilized oligonucleotide strands and sufficient enough as for hybridization to occur, thereby producing hybridized oligonucleotide material;

e) removing said oligonucleotide strands that did not hybridize to said immobilized oligonucleotide strands;

f) recovering said hybridized oligonucleotide material from said sampling medium;

g) transferring a small amount of said recovered hybridized oligonucleotide material in an amount sufficient enough as to provide a detectable quantity of hybridized oligonucleotide material to a SERS active substrate; and

h) analyzing said SERS active substrate containing said hybridized oligonucleotide material.

2. The method of claim 1 wherein said oligonucleotide strand comprises a strand of Deoxyribonucleic acid, Ribonucleic acid or Peptide nucleic acid.

3. The method of claim 1 wherein said sample medium is a nitrocellulose filter.

4. The method of claim 1 wherein said sample medium are microparticulates.

5. The method of claim 4 wherein said microparticulates have magnetic characteristics.

6. The method of claim 1 wherein steps (f) and (g) comprise a blotting procedure.

7. The method of claim 1 wherein said SER gene probe comprises two oligonucleotide strands having at least one SERS label intercalated between said oligonucleotide strands.