Piezoelectric device for detection of polynucleotide hybridization

• US 5,595,908 A
• Filed: 08/11/1989
• Issued: 01/21/1997
• Est. Priority Date: 09/26/1985
• Status: Expired due to Term

• Current Assignee: University of Southern Mississippi
• Original Assignee: University of Southern Mississippi
• Inventors: Evans, Jeffrey A., Fawcett, Newton C.
• Primary Examiner(s): Zitomer, Stephanie W.
• Assistant Examiner(s): Sisson, Bradley L.
• Application Number:
  US07/392,734
• Time in Patent Office: 2,720 Days
• Field of Search: 436/500, 310/316, 310/311, 310/312, 329/117, 422/69, 422/70, 422/55-57, 422/52, 422/50, 935/111, 935/86, 435/6, 435/287, 435/291
• US Class Current: 435/287.2
• CPC Class Codes: C12Q 1/6825 Nucleic acid detection involving sensors
• CPC Class Codes: C12Q 2565/607 being a sensor, e.g. electrode
• CPC Class Codes: G01N 2291/0256 Adsorption, desorption, surface mass change, e.g. on biosensors
• CPC Class Codes: G01N 2291/0257 with a layer containing at least one organic compound
• CPC Class Codes: G01N 29/036 by measuring frequency or resonance of acoustic waves
• CPC Class Codes: G01N 33/542 with steric inhibition or signal modification, e.g. fluorescent quenching
• CPC Class Codes: G01N 33/54373 involving physiochemical end-point determination, e.g. wave-guides, FETS, gratings

Abstract

A system for detecting polynucleotide hybridization is disclosed. A polynucleotide is immobilized on a surface of a piezoelectric crystal. The resonance frequency of the piezoelectric crystal is measured through means for determining the resonant frequency of a piezoelectric crystal. A separate source of polynucleotide is exposed to the polynucleotide-coated piezoelectric crystal for a sufficient length of time and under conditions suitable for hybridization. The resonance frequency of the crystal is then again measured, and the difference between the resonance frequency before and after the incubation period will indicate the extent of hybridization.

Claims

1. A device for detecting polynucleotide hybridization comprising:

(a) a piezoelectric crystal;

(b) a polynucleotide attached to a surface of said crystal by a linker; and

(c) a means for determining the resonance frequency of said crystal.

2. The device of claim 1 wherein said linker is selected from the group comprising organic compounds characterized by the presence of an azido group.

3. The device of claim 1 wherein said linker is selected from 2,6-bis(4-azidobenzylidene) cyclohexanone and triazido trinitrobenzene.

4. A device for detecting polynucleotide hybridization comprising:

(a) a piezoelectric crystal;

(b) a polynucleotide attached to a surface of said crystal by a polymer coated on at least a portion of said crystal, said polymer being selected from a class of polymers characterized by hydrolyrically stable, hydrophobic backbones substituted with reactive pendant groups; and

(c) a means for determining the resonance frequency of said crystal.

5. The device of claim 4 wherein said polymer is selected from the group of polymers consisting of poly(butyl methacrylate) and polyurethane.

6. The device of claim 4 wherein said polymer is a synthetic polynucleotide.

7. The device of claim 4 wherein said polymer is selected from the group consisting of the copolymers of styrene and p-x-methylstyrene, ethylene and N-(6-x-hexyl)-acrylamide, propylene and N-(6-x-hexy)-acrylamide, alkylated siloxanes and 6-x-hexyl siloxanes and alkyl-substituted siloxanes, where X is a reactive group capable of combining with polynucleotides or derivatized polynucleotides.

8. A device for detecting polynucleotide hybridization comprising:

(a) a piezoelectric crystal;

(b) a polynucleotide attached to a surface of said crystal by enzymatically ligating said polynucleotide using DNA or RNA ligase to a different polynucleotide in order to then covalently bond said different polynucleotide by chemical reaction to a functional group which is in turn chemically bonded to said crystal surface; and

(c) a means for determining the resonance frequency of said crystal.

9. A device for detecting polynucleotide hybridization comprising:

(a) a piezoelectric crystal;

(b) a polynucleotide covalently bonded to a surface of said crystal; and

(c) a means for determining the resonance frequency of said crystal.