Impedance-based bacterial detection system

• US 9,709,516 B2
• Filed: 02/14/2013
• Issued: 07/18/2017
• Est. Priority Date: 02/15/2012
• Status: Active Grant

• Current Assignee: Becton, Dickinson & Co
• Original Assignee: Becton, Dickinson & Co
• Inventors: Berndt, Klaus W.
• Primary Examiner(s): Knight, Teresa E
• Application Number:
  US14/378,547
• Publication Number:
  US 20150005196A1
• Time in Patent Office: 1,615 Days
• CPC Class Codes: C12M 41/36 of biomass, e.g. colony counters or by turbidity measurements electrooptical investigation of individual particles G01N15/14, flow cytometers G01N15/1404
• CPC Class Codes: C12Q 1/04 Determining presence or kind of microorganism; Use of selective media for testing antibiotics or bacteriocides; Compositions containing a chemical indicator therefor C12Q1/6897 takes precedence
• CPC Class Codes: G01N 27/02 by investigating impedance
• CPC Class Codes: G01N 27/026 Dielectric impedance spectroscopy electrochemical impedance spectroscopy for measuring corrosion G01N17/02
• CPC Class Codes: G01N 33/48735 Investigating suspensions of cells, e.g. measuring microbe concentration by chemical means C12Q1/04; colony counters C12M1/34; concentration of particle suspensions in general G01N15/06

Abstract

A method and apparatus for determining the presence or absence of microorganisms in a liquid sample. A vessel with an electrode disposed therein receives a volume of liquid to be tested. A second electrode is also provided, both electrodes in physical contact with the liquid sample. A time varying signal is applied to one electrode, and the other electrode is coupled to a phase sensitive signal detector. The phase sensitive signal detector determines a frequency at which an out of phase signal amplitude is zero. This zero-crossing frequency is used as a baseline, and changes in the zero-crossing frequency are an indication of microbial growth.

Claims

1. An impedance-based microbial growth detection method comprising the steps of:

providing at least one vessel containing a liquid sample suspected of containing microorganisms disposed between and in contact with a first electrode and a second electrode;

providing a time-varying electrical signal to the first electrode in contact with the liquid sample, wherein the second electrode is electrically connected to a phase-sensitive signal detector; and

selecting a frequency of said time-varying electrical signal so that an out-of-phase signal amplitude measured by said phase-sensitive signal detector becomes equal to zero at the selected frequency;

measuring the out-of-phase signal amplitude over time with the phase-sensitive signal detector at the selected frequency, wherein an increase in the signal amplitude over time at the selected frequency indicates microbial growth within the liquid sample; and

after measuring, detecting the presence or absence of microbial growth.

2. The method of claim 1 wherein the at least one vessel is an array of vessels at least a plurality of which contain the liquid sample suspected of containing microorganisms disposed between and in contact with the first electrode and the second electrode disposed on the array of vessels, the method further comprising:

providing a time-varying electrical signal generated to the first electrode of the plurality of vessels in the array of vessels, and

receiving the time-varying signal transmitted through the plurality of vessels; and

transmitting the signal to the phase-sensitive signal detector.

3. The method of claim 2 wherein the array of vessels share a common head space.

4. The method of claim 1 wherein the volume of the sample is about 40 mL.

5. The method of claim 1 wherein the sample is combined with growth media in which microorganisms, if present in the sample, will grow.

6. The method of claim 5 wherein the sample is about 10 mL of blood combined with about 30 mL of growth media.

7. The method of claim 1 wherein the first electrode is a bottom electrode that forms the bottom of the vessel and the second electrode is a top electrode extending into the vessel to contact the sample.

8. An impedance-based microbial growth detection method comprising the steps of:

providing a sample container having at least one vessel containing a liquid sample suspected of containing microorganisms disposed between and in contact with a first electrode and a second electrode;

providing a time-varying electrical signal to the first electrode in contact with the liquid sample, wherein the second electrode is electrically connected to a phase-sensitive signal detector; and

measuring an out-of-phase signal amplitude over time with the phase-sensitive signal detector,

determining a frequency at which the out of phase amplitude is zero by tuning the frequency of said electrical signal so that the out-of-phase signal amplitude measured by said detector becomes equal to zero;

repeating the determining step at predetermined time intervals; and

after measuring and repeated determining, detecting the presence or absence of microbial growth,

wherein an increase in said frequency over time is an indication of microbial growth within said liquid sample.

9. The method of claim 8 wherein the at least one vessel is an array of vessels at least a plurality of which contain the liquid sample suspected of containing microorganisms disposed between and in contact with the first electrode and the second electrode disposed on the array of vessels, and the impedance-based detection method further comprising:

providing a time-varying electrical signal generated to the first electrode of the plurality of vessels in the array of vessels, and

receiving the time-varying signal transmitted through the plurality of vessels; and

transmitting the time-varying signal to the phase-sensitive signal detector.

10. The method of claim 9 wherein the array of vessels share a common head space.

11. The method of claim 9 wherein the first electrode is a bottom electrode that forms the bottom of the vessels and the second electrode is a top electrode extending into the array of vessels to contact the sample.

12. The method of claim 8 wherein the volume of the sample is about 40 mL.

13. The method of claim 8 wherein the sample is combined with growth media in which microorganisms, if present in the sample, will grow.

14. The method of claim 13 wherein the sample is about 10 mL of blood combined with about 30 mL of growth media.