Apparatus and method for detection of microorganisms

• US 5,164,796 A
• Filed: 12/01/1991
• Issued: 11/17/1992
• Est. Priority Date: 03/15/1988
• Status: Expired due to Term

• Current Assignee: Biomerieux Incorporated (Compagnie Merieux Alliance SAS)
• Original Assignee: Akzo Nobel N.V. (Nouryon BV)
• Inventors: Di Guiseppi, James L., Thorpe, Thurman C.
• Primary Examiner(s): McGraw, Vincent P.
• Assistant Examiner(s): KEESEE, LACHARLES
• Application Number:
  US07/649,147
• Time in Patent Office: 352 Days
• Field of Search: 356/345, 356/346, 356/412, 356/445, 356/416, 356/39, 250/341, 435/84, 435/291, 422/68
• US Class Current: 356/445
• 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: G01N 21/253 for batch operation, i.e. multisample apparatus analytical automats G01N35/00

Abstract

An instrument for monitoring the color change of an indicator element sealed within a sterile vessel and exhibiting a color change in response to a change in the pH of CO₂ level within the sterile vessel as a result of microbacterial growth within a specimen contained in the sterile vessel. The present invention includes means for continuously monitoring the absolute value of the pH and or CO₂ level within the vessel and or monitoring the rate of change of the absolute value of the pH level.

Claims

1. An instrument for monitoring microbial growth in a specimen, comprising:

a sealable, sterilizable container having an internal chamber in which the specimen is cultured in a sterile culture medium, the container having at least one transparent section;

a sterilizable indicator means located in the container in the region of the transparent section, said indicator means exhibiting a change in its measurable properties detectable through said transparent section upon exposure to metabolites of microbial growth, whereby changes in the indicator means can be monitored from the exterior of the container through said transparent section, thereby monitoring microbial growth without entering the container after sealing;

an emitter means for emitting an emitter signal that interacts with at least one measurable property of said indicator means, whereby an indicator signal is produced, positioned relative to said indicator means so that said emitter signal strikes said indicator means through the transparent section;

a detector means positioned relative to said indicator means for receiving the indicator signal from said indicator means through the transparent section and for producing a detector signal corresponding thereto;

processing means for receiving said detector signal and for processing said detector signal to evaluate changes in or the magnitude of the measurable property of said indicator means and thereby monitor microbial growth in said sealable container after said container has been sealed.

2. An instrument according to claim 1, wherein said processing means for receiving said detector signal is a circuit means.

3. An instrument according to claim 1, said emitter means including a light emitting diode.

4. An instrument according to claim 3, said detector means including a photodiode receptive to the indicator signal produced as a result of the emission of said light emitting diode.

5. An instrument according to claim 3, said processing means for receiving said detector signal including a stabilized current source for said light emitting diode.

6. An instrument according to claim 2, said circuit means including:

computing means for receiving said detector signal at selected time intervals and for comparing any changes in the characteristics of said detector signal during each of said time intervals.

7. An instrument according to claim 2, said circuit means including computation means for periodic sampling of said detector signal, for comparing samples of said detector signal, and for calculating the rate of change of said samples.

8. An instrument according to claim 1 further comprising means for detecting a rate of change of a measurable property of said indicator means and for measuring the duration of said rate of change.

9. An instrument according to claim 1, wherein the indicator means comprises a membrane and an indicator medium, the indicator medium exhibiting a detectable change when exposed to microbial metabolites.

10. An instrument according to claim 9, wherein the membrane is secured to an interior wall of the container.

11. An instrument according to claim 1, wherein the indicator means is disposed against an interior wall of the container and the indicator means comprises a membrane that separates the indicator means from the culture medium.

12. An instrument set forth in claim 1, wherein the metabolite is CO₂.

13. An instrument according to claim 1, wherein the indicator means comprises a chemical responsive to pH.

14. The instrument set forth in claim 1, wherein said measurable property is selected from the group consisting of light absorbance, phosphorescence, light scattering, refraction, fluorescence, and light reflectance characteristics.

15. An instrument according to claim 8, further comprising means for detecting acceleration of change of a measurable property of the indicator means.

16. An instrument according to claim 15, also measuring the duration of said acceleration.

17. An instrument according to claim 1, further comprising means for measuring the magnitude of said detector signal.

18. An instrument according to claim 17, also measuring the duration of the magnitude of said detector signal.

24. A method of determining the presence of microbes in a specimen, comprising the steps of:

providing the instrument of claim 1;

introducing the specimen into the sterile container;

incubating the specimen in the container;

monitoring for a change in a measurable property of the indicator means with the detector means at at least one time interval;

calculating the rate of change of the measurable property, whereby a first derivative of the property is determined; and

analyzing the magnitude of the first derivative of the measurable property to establish the presence of microbes in the specimen.

25. A method according to claim 24, including the further steps of:

calculating the second derivative of the measurable property; and

analyzing the magnitude of the second derivative of the measurable property whereby the presence of microbes in the specimen is ascertained.

26. A method according to claim 25, including the further steps of:

measuring the duration of said second derivative.

27. The method set forth in claim 24, wherein said measurable property is selected from the group consisting of light absorbance, phosphorescence, light scattering, refraction, fluorescence, and light reflectance characteristics.

28. A method of determining the presence of microbes in a specimen, comprising the steps of:

providing the instrument of claim 1;

introducing the specimen into the sterile container;

incubating the specimen in the container;

monitoring for a change in a measurable property of the indicator means with the detector means at at least one time interval; and

measuring the magnitude of the measurable property to establish the presence of microbes in the specimen.

19. A method for detecting microbial growth in a specimen, comprising the steps of:

providing a sterilizable container, the container having at least one transparent section in a wall thereof and an indicator means that changes when exposed to metabolites of microbial growth disposed in the container in the region of the transparent section;

providing an emitter means for emitting an emitter signal that interacts with at least one measurable property of said indicator means, whereby an indicator signal is produced, and positioned relative to said indicator means so that said emitter signal strikes said indicator means through the transparent section;

providing a detector means positioned relative to said indicator means for receiving the indicator signal from said indicator means and for producing a detector signal corresponding thereto;

providing a processing means for receiving said detector signal and for processing said detector signal to evaluate the change in or the magnitude of the measurable properties of said indicator means;

introducing the specimen under sterile conditions into the container;

incubating the specimen in the container; and

detecting a change in or magnitude of the detector signal, thereby detecting microbial growth in the sealable bottle after the container has been sealed.

20. A method according to claim 19, comprising the further steps of:

receiving said detector signal at selected time intervals; and

comparing any changes in the characteristics of said detector signal during said time intervals.

21. A method according to claim 19, including the further steps of:

sampling said detector signal;

comparing successive samples of said detector signal; and

calculating the rate of increase in the rate of change between successive samples.

22. A method according to claim 19, including the further steps of:

detecting a rate of change in the detector signal; and

measuring the duration of said rate.

23. The method set forth in claim 19, wherein said measurable property is selected from the group consisting of light absorbance, phosphorescence, light scattering, refraction, fluorescence, and light reflectance characteristics.