WO1999045095A1 - Disposable apparatus for determining antibiotic sensitivity of bacteria - Google Patents
Disposable apparatus for determining antibiotic sensitivity of bacteria Download PDFInfo
- Publication number
- WO1999045095A1 WO1999045095A1 PCT/US1999/004511 US9904511W WO9945095A1 WO 1999045095 A1 WO1999045095 A1 WO 1999045095A1 US 9904511 W US9904511 W US 9904511W WO 9945095 A1 WO9945095 A1 WO 9945095A1
- Authority
- WO
- WIPO (PCT)
- Prior art keywords
- reagent
- growth medium
- antibiotic
- sensible
- target microorganism
- Prior art date
Links
Classifications
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
- C12Q1/02—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving viable microorganisms
- C12Q1/18—Testing for antimicrobial activity of a material
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
- C12M23/00—Constructional details, e.g. recesses, hinges
- C12M23/42—Integrated assemblies, e.g. cassettes or cartridges
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
- C12M41/00—Means for regulation, monitoring, measurement or control, e.g. flow regulation
- C12M41/46—Means for regulation, monitoring, measurement or control, e.g. flow regulation of cellular or enzymatic activity or functionality, e.g. cell viability
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
- C12M23/00—Constructional details, e.g. recesses, hinges
- C12M23/02—Form or structure of the vessel
- C12M23/12—Well or multiwell plates
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
- C12M25/00—Means for supporting, enclosing or fixing the microorganisms, e.g. immunocoatings
- C12M25/06—Plates; Walls; Drawers; Multilayer plates
Definitions
- the present invention relates to apparatus for determining a microorganism's sensitivity to an antibiotic in general, and to apparatus for determining the minimum inhibitory concentration of an antibiotic relative to a microorganism in particular.
- the determination of the minimum inhibitory concentration (MIC) of an antibiotic is an essential laboratory test to determine the sensitivity of a microorganism, usually a bacterium, to specific antibiotics.
- the MIC refers to the minimum concentration of an antibiotic necessary to prevent the microorganism from growing.
- the type and dose of antibiotics is often predicated upon this type of test, making rapid and accurate results critical to both patient care and cost-effective treatment.
- Antibiotic sensitivity testing is most commonly performed using the qualitative Kirby- Bauer plate method, but for a quantitative MIC analysis, the tube dilution method is most commonly used.
- the Kirby-Bauer test utilizes a plate covered with a uniform layer of microbiological growth medium specifically formulated for the test at hand. A number of disks are placed on the layer of growth medium, each containing a specific concentration of an antibiotic being evaluated. Bacteria grows on the medium forming a visible coating, except in the area (generally referred to as the "clear zone") around those disks having sufficient antibiotic concentration to inhibit bacterial growth. The size of the clear zone surrounding a disk is indicative of the organism's sensitivity to the antibiotic contained in that particular disk; i.e., the larger the clear zone, the greater the organism's sensitivity to the antibiotic contained in the disk.
- the Kirby-Bauer test is popular because of its simplicity and its ability to evaluate multiple antibiotics at once.
- a disadvantage of the Kirby-Bauer test is that there are a number of variables which affect the antibiotic concentration at any given point in the growth medium, and thus do not allow a MIC to be calculated. Formulae have been published for calculating the approximate MIC based upon the clear zone size, but these formulae are rarely used and are considered to be approximations at best.
- United States Patent No. 5,702,684 discloses a method for monitoring antibiotic levels for determining when the antibiotics should be replenished in an industrial plumbing system using a fluorescent marker. This method, however, does not allow the determination of a MIC or any type of antibiotic sensitivity measurement. What is needed is an apparatus for determining the MIC of an antibiotic for a target microorganism, one that can determine the MIC in a minimum amount of time, one that provides an accurate MIC, one that can simultaneously determine the MIC's of several antibiotics for a target microorganism, and one that is cost effective.
- an apparatus for testing the minimum inhibitory concentration of antibiotic for a target microorganism includes a body, a sheet of microorganism growth medium, and a sensible reagent. The body includes a well for receiving the sheet of microorganism growth medium.
- An advantage of the present invention is that an apparatus for determining the MIC of an antibiotic for a target microorganism is provided that gives accurate results in a minimum amount of time.
- the present invention uses a sensible reagent including a marker having a signal proportional to its concentration which, in turn, is proportional to the concentration of the antibiotic.
- the MIC of antibiotic at the growth boundary can be determined by sensing the marker signal at the growth boundary. Accordingly, the exact MIC of antibiotic can be determined rather than an approximation, and can be determined without a multitude of time consuming dilution steps.
- Another advantage of the present invention is that the MIC's of several antibiotics for a target microorganism can be determined concurrently using the present invention.
- Another advantage of the present invention is that a cost effective apparatus for determining the MIC of an antibiotic for a target microorganism is provided.
- the ability of the present invention method to provide accurate MIC information obviates the need for multiple expensive antibiotic dilutions as are required in the tube dilution method.
- a person of skill in the art will recognize that minimizing expensive medical laboratory time and laboratory assets make the present invention method considerably less expensive than presently available methods.
- Another advantage of the present invention is that a "user-friendly" apparatus for determining the MIC of an antibiotic for a target microorganism is provided.
- the present invention cassette facilitates the MIC testing, particularly multiple MIC tests, minimizes the opportunity for sample spillage, and can be readily disposed of after the test. These qualities and others make the present invention cassette attractive as a disposable.
- FIG.l is a diagrammatic view of the present invention cassette.
- FIG.2 is a width-wise cross-sectional view of the cassette shown in FIG.l.
- FIG.3 is a cross-sectional view similar to that shown in FIG.2, illustrating a cassette embodiment having a plurality of wells.
- FIG.4 is a length- wise cross sectional view of the cassette shown in FIG.1.
- the transparent well cap 22 attaches to the body 12 above the well 14 to protect and maintain the growth medium 16 and absorbent strips 20 inside the well 14.
- the body 12 includes a plurality of wells 14, each similar to that described above.
- a port 30 (FIG.1) through which a target microorganism solution can be distributed to the growth medium 16 is disposed in the well cap 22.
- the port 30 may alternatively be disposed in the body 12 of the cassette 10, or be formed between the body 12 and the well cap 22.
- the cassette 10 further includes a machine readable information label 32 and a user readable information label 34.
- the machine readable information label 32 includes a data block 36 containing pertinent information such as the test to be performed, calibration constants, patient identification, or the like, in a machine readable format such as a bar code or magnetic strip.
- the machine readable label 32 may directly contain all of the information necessary to enable an analytical device 61 to perform the analysis at hand.
- the machine readable label 32 may instruct the analytical device 61 to access data files contained within the analytical device or remotely accessed by the analytical device.
- the label 32 directly or indirectly contains the information necessary to enable an analytical device 61 to perform the analysis at hand.
- the sheet of growth medium 16 is preferably a hydroscopic material such as polyvinyl alcohol, hydroxyethyl starch, phytagel, modified agar, or the like, which is admixed with materials capable of sustaining growth of the target microorganism and may also contain agents which serve to highlight microbial growth. Examples of such agents include general growth indicators such as disclosed in United States Patent Nos. 4,049,099 and 5,501,959. Dehydrated growth mediums 16 that may be rehydrated during use are favored because they can be readily stored for extended periods of time.
- the sheet of growth medium 16 has a uniform thickness "T" (see FIGS. 2-5) in the areas 40, referred to as the "active" areas 40, where the sensible reagent 18 is incorporated.
- the active areas 40 of the growth medium sheet 16 are separated by trenches 42, thereby forming rows of active areas 40. Dividers 44 can be attached to the well cap 22 to help separate adjacent active areas 40.
- a fluorescent marker having excitation or emission wavelengths outside the range of the excitation or emission wavelengths of the growth medium, and one that does not bind to the growth medium 16 or the target microorganism may be used.
- the sensible reagent 18 may be directly or indirectly incorporated into the growth medium 16. Direct incorporation may be accomplished by several methods including, but not limited to, injecting the sensible reagent 18 into the growth medium 16, or applying the reagent 18 onto a surface of the growth medium 16 and allowing the reagent 18 to diffuse into the growth medium 16. Injecting the sensible reagent 18 into the growth medium 16 may be advantageous for a reagent 18 with a low diffusion rate (once in the growth medium 16, the reagent 18 can diffuse locally to form a desirable concentration gradient).
- the strip 48 has a first quantity of sensible reagent 18 which, when incorporated into the growth medium 16, will create an antibiotic concentration sufficient to inhibit growth of the target microorganism within the growth medium 16.
- the strip 48 has a second quantity of sensible reagent which, when incorporated into the growth medium 16, will create an antibiotic concentration insufficient to inhibit growth of the target microorganism within the growth medium 16.
- the first quantity of reagent 18 is greater than the second quantity, and a gradient 49 of decreasing reagent quantities extends between the first and second ends 52,54.
- the section of growth medium 16 having detectable target microorganism growth will be contiguous with a section of growth medium 16 having substantially no detectable target microorganism growth.
- the border between the two sections is referred to as the growth boundary.
- the section of growth medium 16 having detectable growth of target microorganism is that in which the target microorganism is substantially uninhibited by the antibiotic.
- the section having no detectable growth is that in which the microorganism is substantially inhibited by the antibiotic.
- the growth boundary coincides with the MIC of the antibiotic for the target microorganism being evaluated.
Landscapes
- Chemical & Material Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Health & Medical Sciences (AREA)
- Organic Chemistry (AREA)
- Wood Science & Technology (AREA)
- Engineering & Computer Science (AREA)
- Zoology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- General Health & Medical Sciences (AREA)
- Microbiology (AREA)
- Biotechnology (AREA)
- Biochemistry (AREA)
- General Engineering & Computer Science (AREA)
- Genetics & Genomics (AREA)
- Analytical Chemistry (AREA)
- Biomedical Technology (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Sustainable Development (AREA)
- Cell Biology (AREA)
- Toxicology (AREA)
- Physics & Mathematics (AREA)
- Biophysics (AREA)
- Clinical Laboratory Science (AREA)
- Immunology (AREA)
- Molecular Biology (AREA)
- Apparatus Associated With Microorganisms And Enzymes (AREA)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
- Investigating Or Analysing Biological Materials (AREA)
Abstract
Description
Claims
Priority Applications (5)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
JP2000534627A JP2002516660A (en) | 1998-03-07 | 1999-03-02 | A disposable device for determining bacterial antibiotic susceptibility. |
CA002321686A CA2321686A1 (en) | 1998-03-07 | 1999-03-02 | Disposable apparatus for determining antibiotic sensitivity of bacteria |
AU27996/99A AU743628B2 (en) | 1998-03-07 | 1999-03-02 | Disposable apparatus for determining antibiotic sensitivity of bacteria |
EP99908609A EP1064352A4 (en) | 1998-03-07 | 1999-03-02 | Disposable apparatus for determining antibiotic sensitivity of bacteria |
NO20004450A NO20004450L (en) | 1998-03-07 | 2000-09-06 | Disposable device for determining the sensitivity of bacteria to antibiotics |
Applications Claiming Priority (4)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
US7721698P | 1998-03-07 | 1998-03-07 | |
US60/077,216 | 1999-02-23 | ||
US09/256,451 | 1999-02-23 | ||
US09/256,451 US6022734A (en) | 1998-03-07 | 1999-02-23 | Disposable apparatus for determining antibiotic sensitivity of bacteria |
Publications (1)
Publication Number | Publication Date |
---|---|
WO1999045095A1 true WO1999045095A1 (en) | 1999-09-10 |
Family
ID=26759033
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
PCT/US1999/004511 WO1999045095A1 (en) | 1998-03-07 | 1999-03-02 | Disposable apparatus for determining antibiotic sensitivity of bacteria |
Country Status (8)
Country | Link |
---|---|
US (1) | US6022734A (en) |
EP (1) | EP1064352A4 (en) |
JP (1) | JP2002516660A (en) |
CN (1) | CN1195842C (en) |
AU (1) | AU743628B2 (en) |
CA (1) | CA2321686A1 (en) |
NO (1) | NO20004450L (en) |
WO (1) | WO1999045095A1 (en) |
Cited By (5)
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---|---|---|---|---|
EP1120467A2 (en) * | 2000-01-05 | 2001-08-01 | Stephen Clark Wardlaw | Method and apparatus for determining the sensitivity of a microorganism to a growth-altering agent |
DE102013225037A1 (en) * | 2013-12-05 | 2015-06-11 | Fraunhofer-Gesellschaft zur Förderung der angewandten Forschung e.V. | Method for detecting resistant germs and device for carrying out the same |
WO2017006055A1 (en) * | 2015-07-09 | 2017-01-12 | bioMérieux | Method for detecting a presence or absence of at least a first zone of inhibition |
EP3725871A4 (en) * | 2017-12-13 | 2021-09-29 | Hitachi High-Tech Corporation | Bacterial test plate having antibacterial agent introduced thereinto, and transparent plate |
WO2023118904A1 (en) * | 2021-12-23 | 2023-06-29 | The University Of Liverpool | Hydrogels and uses thereof |
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US6929953B1 (en) | 1998-03-07 | 2005-08-16 | Robert A. Levine | Apparatus for analyzing biologic fluids |
US6723290B1 (en) * | 1998-03-07 | 2004-04-20 | Levine Robert A | Container for holding biologic fluid for analysis |
US6472166B1 (en) * | 2000-02-17 | 2002-10-29 | Wardlaw Partners Lp | Method for determining the effects of a growth-altering agent on a microbial colony |
US20020136732A1 (en) * | 2000-04-23 | 2002-09-26 | Houston L. L. | Compositions comprising carriers and transportable complexes |
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US20030166160A1 (en) * | 2001-09-06 | 2003-09-04 | Hawley Stephen B. | Compounds and molecular complexes comprising multiple binding regions directed to transcytotic ligands |
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-
1999
- 1999-02-23 US US09/256,451 patent/US6022734A/en not_active Expired - Fee Related
- 1999-03-02 CA CA002321686A patent/CA2321686A1/en not_active Abandoned
- 1999-03-02 JP JP2000534627A patent/JP2002516660A/en active Pending
- 1999-03-02 WO PCT/US1999/004511 patent/WO1999045095A1/en active IP Right Grant
- 1999-03-02 EP EP99908609A patent/EP1064352A4/en not_active Withdrawn
- 1999-03-02 AU AU27996/99A patent/AU743628B2/en not_active Ceased
- 1999-03-02 CN CNB998037419A patent/CN1195842C/en not_active Expired - Fee Related
-
2000
- 2000-09-06 NO NO20004450A patent/NO20004450L/en not_active Application Discontinuation
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Non-Patent Citations (1)
Title |
---|
See also references of EP1064352A4 * |
Cited By (12)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
EP1120467A2 (en) * | 2000-01-05 | 2001-08-01 | Stephen Clark Wardlaw | Method and apparatus for determining the sensitivity of a microorganism to a growth-altering agent |
SG99326A1 (en) * | 2000-01-05 | 2003-10-27 | Stephen C Wardlaw | Method and apparatus for determining the sensitivity of a microorganism to a growth altering agent |
EP1120467A3 (en) * | 2000-01-05 | 2003-11-19 | Stephen Clark Wardlaw | Method and apparatus for determining the sensitivity of a microorganism to a growth-altering agent |
DE102013225037A1 (en) * | 2013-12-05 | 2015-06-11 | Fraunhofer-Gesellschaft zur Förderung der angewandten Forschung e.V. | Method for detecting resistant germs and device for carrying out the same |
DE102013225037B4 (en) * | 2013-12-05 | 2016-08-25 | Asklepios Kliniken Verwaltungsgesellschaft mbH | Method for detecting resistant germs and device for carrying out the same |
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FR3038620A1 (en) * | 2015-07-09 | 2017-01-13 | Biomerieux Sa | METHOD FOR DETECTING A PRESENCE OR ABSENCE OF AT LEAST ONE FIRST INHIBITION AREA |
US10870876B2 (en) | 2015-07-09 | 2020-12-22 | Biomerieux | Method for detecting a presence or absence of at least one first zone of inhibition |
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WO2023118904A1 (en) * | 2021-12-23 | 2023-06-29 | The University Of Liverpool | Hydrogels and uses thereof |
Also Published As
Publication number | Publication date |
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US6022734A (en) | 2000-02-08 |
EP1064352A1 (en) | 2001-01-03 |
EP1064352A4 (en) | 2006-05-10 |
NO20004450L (en) | 2000-10-23 |
AU2799699A (en) | 1999-09-20 |
CA2321686A1 (en) | 1999-09-10 |
CN1301296A (en) | 2001-06-27 |
AU743628B2 (en) | 2002-01-31 |
CN1195842C (en) | 2005-04-06 |
JP2002516660A (en) | 2002-06-11 |
NO20004450D0 (en) | 2000-09-06 |
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