Apparatus and method for detecting and identifying infectious agents

• US 6,649,357 B2
• Filed: 04/19/2002
• Issued: 11/18/2003
• Est. Priority Date: 12/12/1996
• Status: Expired due to Term

• Current Assignee: Prolume Limited
• Original Assignee: Prolume Limited
• Inventors: Murphy, Randall B., Gaalema, Stephen, Bryan, Bruce J.
• Primary Examiner(s): Le, Long V.
• Assistant Examiner(s): Padmanabhan, Kartic
• Application Number:
  US10/126,798
• Publication Number:
  US 20030059798A1
• Time in Patent Office: 578 Days
• Field of Search: 435/6, 435/7.1, 435/288.7, 435/808, 435/973, 435/975, 435/7.9, 436/164, 436/172, 436/516, 436/527, 436/532, 436/805, 436/524, 422/57, 422/58, 422/68.1, 422/82.05, 422/82.08, 356/215, 356/222, 356/317
• US Class Current: 506/9
• CPC Class Codes: G01N 21/253 for batch operation, i.e. multisample apparatus analytical automats G01N35/00
• CPC Class Codes: G01N 21/763 Bioluminescence
• CPC Class Codes: G01N 33/54366 Apparatus specially adapted for solid-phase testing
• CPC Class Codes: G01N 33/582 with fluorescent label
• CPC Class Codes: Y10S 435/808 Optical sensing apparatus
• CPC Class Codes: Y10S 435/973 Simultaneous determination of more than one analyte
• CPC Class Codes: Y10S 435/975 Kit
• CPC Class Codes: Y10S 436/805 Optical property
• CPC Class Codes: Y10T 436/186 Sulfur dioxide

Abstract

Solid phase methods for the identification of an analyte in a biological medium, such as a body fluid, using bioluminescence are provided. A chip designed for performing the method and detecting the bioluminescence is also provided. Methods employing biomineralization for depositing silicon on a matrix support are also provided. A synthetic synapse is also provided.

Claims

1. A method of detecting and identifying analytes in a biological sample, comprising the steps of:

(a) providing a microelectronic device comprising;

a substrate;

a plurality of micro-locations defined on the substrate, wherein each micro-location is for linking a macromolecule;

an independent photodetector integrated at each micro-location and optically coupled to each micro-location, each photodetector being configured to generate a sensed signal responsive to the photons of light emitted at the corresponding micro-location when a light-emitting chemical reaction occurs at that micro-location, each photodetector being independent from the photodetectors optically coupled to the other micro-locations; and

an electronic circuit coupled to each photodetector and configured to read the sensed signal generated by each photodetector and to generate output data signals therefrom that are indicative of the light emitted at each micro-location by the light-emitting chemical reactions, whereby the device detects photons of light emitted by light-emitting chemical reactions, wherein each micro-location is defined by a portion of the surface of the device;

(b) attaching a macromolecule or plurality of different macromolecules to the substrate at each micro-location on the device, wherein the macromolecule is specific for binding to a selected analyte present in the biological sample;

(c) contacting the sample with the surface of the microelectronic device, whereby any of the selected analytes that are present in the sample bind to the macromolecule attached to the surface at each micro-location;

(d) exposing the surface of the microelectronic device to a second macromolecule or plurality thereof which bind to the selected analyte already bound to the first macromolecule at each micro-location, wherein the second macromolecule comprises a component of a bioluminescence generating reaction;

(e) initiating the bioluminescence generating reaction by contacting the surface of the device with the remaining components of the bioluminescence generation reaction; and

(f) detecting photons of light emitted by the bio-luminescent reaction using the photodetector optically coupled to each micro-location, each photodetector generating a sensed signal representative of the bioluminescence generation at the respective micro-location.

2. The method of claim 1, further comprising reading the sensed signal generated by each photodetector and generating output data signals therefrom that are indicative of the bioluminescence emitted at each microlocation due to a luciferase-luciferin reaction.

3. The method of claim 1, between the contacting and exposing steps, further comprising, washing the sample from the surface of the microelectronic device after waiting a sufficient period of time for the selected analytes present in the sample to bind to the macromolecule attached to the surface at each micro-location.

4. The method of claim 1, wherein the macromolecules are antibodies.

5. The method of claim 1, wherein the attaching step includes attaching a plurality of different receptor antibodies to a plurality of different micro-locations, each of the different antibodies being specific to bind a different selected analyte present in the biological sample.

6. The method of claim 1, wherein the attaching step includes immersing the microelectronic device into a fluid volume of the biological sample.

7. The method of claim 1, wherein the analytes that are detected or identified are infectious agents.

8. The method of claim 1, wherein the bioluminescence generating system further comprises a fluorescent protein.

9. The method of claim 8, wherein the fluorescent protein is selected from the group consisting of green fluorescent protein (GFP), blue fluorescent protein (BFP) and a phycobiliprotein.

10. The method of claim 1, wherein the component of the bioluminescence generating system is a luciferase or luciferin.

11. The method of claim 10, wherein the luciferase is a photoprotein.

12. The method of claim 11, wherein the bioluminescence generating system is selected from the group consisting of the Aequorea, Vargula, Renilla, Obelin, Porichthys, Odontosyllis, Aristostomias, Pachystomias, firefly, and bacterial systems.