Method and apparatus for forming of an automated sampling device for the detection of;utilizing an electrochemical aptamer biosensor

• US 9,310,363 B2
• Filed: 04/21/2010
• Issued: 04/12/2016
• Est. Priority Date: 01/07/2010
• Status: Active Grant

• Current Assignee: Sensor-Kinesis Corporation
• Original Assignee: Sensor-Kinesis Corporation
• Inventors: Shachar, Yehoshua, Wu, Winston, Chen, Thomas, Farkas, Leslie, Jordan, Brett, Luboff, Paladin, Chan, Herwin, Zimmerman, Kyle
• Primary Examiner(s): Zara, Jane
• Application Number:
  US12/764,828
• Publication Number:
  US 20110162979A1
• Time in Patent Office: 2,183 Days
• Field of Search: 506/13, 424/9.1, 435/6.11, 435/91.1, 435/91.31, 204/403.01, 205/777.5, 536/23.1
• US Class Current: 1/1
• CPC Class Codes: G01N 2333/255 Salmonella (G)
• CPC Class Codes: G01N 33/5438 Electrodes
• CPC Class Codes: G01N 33/56916 Enterobacteria, e.g. shigella, salmonella, klebsiella, serratia

Abstract

An aptamer-based solid-state electrochemical biosensor for label-free detection of Salmonella enterica serovars utilizing immobilized aptamers. The device is realized by forming a matrix array of parallel capacitors, thus allowing the realization of low-cost, portable, fully integrated devices. Protein-aptamer binding modulates the threshold voltage of a circuit, changing the impedance (capacitance) of the circuit. This circuit is further characterized by an electrode coded with a p-Si substrate, enhancing the affinity between the Salmonella outer membrane proteins (OMPs) and the aptamer. An aptamer embedded detection plate is configured within a testing lid device that fits a standard, commercially available polymer specimen jar. A sample is mixed with broth for incubation and cultivation of any present Salmonella bacteria to obtain acceptable concentration of the pathogen for testing. The information obtained can then be transmitted by wireless network.

Claims

1. An apparatus for detecting Salmonella enterica bacteria using an electrochemical aptamer-based sensor array comprising:

a sensor base plate;

a capacitive array disposed on the sensor base plate in a plurality of grooved channels;

a plurality of aptamer molecules disposed within the capacitive array, each of the plurality of aptamer molecules capable of binding to an indicator protein on an outer membrane of the Salmonella enterica, and wherein each aptamer molecule is immobilized to the capacitive array utilizing the Self-Assembled Monolayer (SAM) method;

a printed circuit board coupled to the capacitive array, the printed circuit board comprising means of interfacing and controlling the capacitive array, and

wherein the printed circuit board and capacitive array are powered directly or indirectly to an independent power supply through a USB connection;

a resistive heating element coupled to and powered by the printed circuit board;

a delivery system for delivering a fluid for analysis to the capacitive array, wherein the delivery system comprises a piezoelectric pump coupled to the sensor base plate to ensure proper liquid flow over the capacitive array; and

a weatherproof enclosure sized and configured to house the apparatus, wherein the enclosure comprises a plurality of attachment brackets comprising means for allowing the enclosure to be coupled to the external surface of a shipping container,

wherein the plurality of attachment brackets of the enclosure further comprises means for the enclosure to fit within a pair of extended corrugations on the external surface of a standard shipping container, the enclosure comprising a profile less than that of the extended corrugations on the external surface of the standard shipping container.

2. The apparatus of claim 1 where the capacitive array disposed on the sensor plate is configured in a plurality of tunnels that are enclosed on four sides and open to liquid flow at either end.

3. The apparatus of claim 1 wherein each of the plurality of aptamer molecules further comprises:

a linker molecule acting as an immobilizer coupled to the aptamer molecule;

an amino-silanization layer coupled to the linker molecule; and

a SiO2 insulator coupled to the amino-silanization layer and to the capacitive array.

4. The apparatus of claim 1 wherein the means of interfacing and controlling the capacitive array by the printed circuit board further comprises means for detecting the change in capacitance in the capacitive array due to the presence of Salmonella enterica bacteria between a pair of the plurality of grooved channels.

5. The apparatus of claim 1 where the delivery system comprises means for providing an unrestricted circulation flow of the fluid through the capacitive array.

6. An apparatus for testing for Salmonella enterica bacteria comprising:

a specimen cup;

a lid comprising an aptamer-based sensor array, the lid being removeably coupled to the specimen cup;

a base station comprising means for interfacing with the lid, wherein the base station comprises;

an electrochemical battery array;

a printed circuit board comprising a CPU and a flash memory;

a liquid crystal display screen;

a USB port comprising means for power acquisition and data communication;

a power port comprising means for connecting to an AC or DC power source ranging from 110 to 240 volts; and

a wireless antenna comprising means for communicating in standard Wi-Fi and cellular phone frequencies;

a weatherproof enclosure sized and configured to house the apparatus including the base station, the lid, and the specimen cup, wherein the enclosure comprises;

a solar photo-electric cell;

an electrochemical battery coupled to the base station and comprising means for collecting power from the solar photo-electric cell and delivering that power to the base station;

an external booster antenna coupled to the base station via the USB port on the base station; and

a plurality of attachment brackets comprising means for allowing the enclosure to be coupled to the external surface of a shipping container; and

a strong permanent magnet coupled to the backside of the enclosure and comprising means for semi-permanent attachment of the enclosure to ferrous surfaces,

wherein the plurality of attachment brackets of the enclosure further comprises means for the enclosure to fit within a pair of extended corrugations on the external surface of a standard shipping container, the enclosure comprising a profile less than that of the extended corrugations on the external surface of the standard shipping container.

7. The apparatus of claim 6 wherein the base station comprising means for interfacing with the lid further comprises the base station being removeably coupled to the aptamer-based sensor array within the lid by means of a USB connection.