EP0498920A2
Expired / Lapsed
EP
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Assay method for detecting the presence of bacteria
Assignee: Vicam LP
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Filing: 1991-05-16
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Granted: —
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Expires: 2011-05-16
Abstract
Abstract An assay method is provided to easily and visually detect the presence of organisms capable of being cultured, such as bacteria, characterized by antibody capture of the organism of interest, incubation of the captured cells to form colonies; removal of material from the colonies with a colony lift membrane; and detection of the colony material on the membrane sheet by use of labeled antibodies.
Inventors
- 👤 Thomas L. Benjamin
- 👤 Joan Chen-Wu
- 👤 Thomsen Hansen
- 👤 Barbara Jackson
- 👤 David Livingstone
- 👤 Steven Tannenbaum
- 👤 Gerald Wogan
Bibliographic Details
Application Number
—
Jurisdiction
EP
Data Source
google_patents
Last Synchronized
2026-11-07 14:55
CPC Classifications
G; G01; G01N; G01N33/00; G01N33/48; G01N33/50; G01N33/53; G01N33/543; G01N33/54313; G01N33/54326; C; C12; C12Q; C12Q1/00; C12Q1/02; C12Q1/04; C12Q1/24; C12Q1/68; C12Q1/6804; G01N33/569; G01N33/56911
Claims Analysis
Total Claims: 22
Claim 1
Independent Claim
103 words
A method for detecting the presence of an organism capable of being cultured which comprises: a) combining a sample to be tested for the presence of a selected organism with a solid support having immobilized thereon antibodies to said selected organism, to thereby capture cells of said organism from said sample; b) culturing said captured organism to form colonies of said organism; c) contacting said colonies with a colony lift membrane whereby some colony material adheres to said membrane; and d) treating said membrane to provide visual evidence of the presence on said membrane of material from said colonies of said selected organism.
Claim 2
Dependent Claim
43 words
The method according to claim 1, wherein said solid support is comprised of a matrix selected from the group consisting of magnetic beads, polyacrylamide beads, agarose beads, polysaccharides, cross-linked dextrans, glass beads, latex beads, glass fiber filters, cellulose nitrate filters and nylon filters.
Claim 3
Independent Claim
19 words
The method according to claims 1 or 2, wherein said colony lift membrane is comprised of nitrocellulose or nylon.
Claim 4
Independent Claim
33 words
The method according to any one of claims 1 to 3, wherein after contacting with said organism colonies, said membrane is treated with a fixing agent to fix said colonies to said membrane.
Claim 5
Independent Claim
63 words
The method according to any one of claims 1 to 4, wherein said membrane with colony material adhered thereto is contacted with detector antibodies which bind to said colony material of said selected organism and the presence of said detector antibody on said membrane is detected to provide visual evidence of the presence on said membrane of colony material from said selected organism.
Claim 6
Dependent Claim
20 words
The method according to claim 5, wherein said label is selected from fluorogenic, radioactive, chemiluminescent, bioluminescent, and enzyme substrate molecules.
Claim 7
Independent Claim
20 words
The method according to any one of claims 1 to 6, wherein said organism is a bacteria, yeast or mold.
Claim 8
Dependent Claim
50 words
The method according to claim 7, wherein said bacteria is selected from Listeria, Campylobacter, Escherichia coli, Salmonella, Clostridia, Shigella, Staphylococci, Vibrio, Yersinia, Plesiomonas strigelloides, Bacilli and Aeromonas; said yeast is selected from Kluyveromyces, Pichia, Saccharomyces, Candida and Rhodotorula and said mold is selected from Byssochlamys, fusarium, Geotrichum, Penicillium and Scopulariopsis.
Claim 9
Dependent Claim
12 words
The method according to claim 8, wherein said selected bacteria is Listeria.
Claim 10
Dependent Claim
15 words
The method according to claim 9, wherein said selected bacteria are pathogenic strains of Listeria.
Claim 11
Independent Claim
29 words
The method according to any one of claims 1 to 10, wherein said culturing is conducted by culturing said solid support and organism captured thereon on a solid medium.
Claim 12
Independent Claim
204 words
A method for detecting the presence of bacteria which comprises: a) combining a sample to be tested for the presence of a selected bacteria with a magnetic solid support having immobilized thereon first antibodies to said selected bacteria, to thereby capture cells of said selected bacteria from said sample; b) exposing to a magnetic field said solid support having bound thereto said first antibodies and captured bacteria cells to thereby separate said magnetic solid support from said sample; c) culturing said magnetic solid support and captured bacteria cells on a solid medium to form colonies of said bacteria; d) contacting said colonies with a colony lift membrane whereby colony material from said colonies adheres to said membrane; e) contacting said membrane and said colony material adhered thereto with second antibodies specific for said selected bacteria, whereby said second antibodies bind to said colony material from said colonies of said selected bacteria; f) contacting said membrane with labeled antibodies specific for said second antibodies, whereby said labeled antibodies bind to said second antibodies bound to said colonies of said selected bacteria; and g) treating said membrane to provide visual evidence of the presence on said membrane of said label and colonies of said selected bacteria.
Claim 13
Dependent Claim
17 words
The method according to claim 12, wherein said colony lift membrane is comprised of nitrocellulose or nylon.
Claim 14
Independent Claim
30 words
The method according to claims 12 or 13, wherein after contacting with said bacteria colonies, said membrane is treated with a fixing agent to fix said colonies to said membrane.
Claim 15
Independent Claim
25 words
The method according to any one of claims 12 to 14, wherein said label is selected from fluorogenic, radioactive, chemiluminescent, bioluminescent, and enzyme substrate molecules.
Claim 16
Independent Claim
32 words
The method according to any one of claims 12 to 15, wherein said bacteria is selected from Listeria, Campylobacter, Escherichia coli, Salmonella, Clostridia, Shigella, Staphylococci, Vibrio, Yersinia, Plesiomonas strigelloides, Bacilli and Aeromonas.
Claim 17
Dependent Claim
12 words
The method according to claim 16, wherein said selected bacteria is Listeria.
Claim 18
Independent Claim
22 words
The method according to any one of claims 12 to 17, wherein said first antibodies are polyclonal antibodies to said selected bacteria.
Claim 19
Independent Claim
25 words
The method according to any one of claims 12 to 18, wherein said second antibodies and labeled antibodies are, or are derived from, monoclonal antibodies.
Claim 20
Independent Claim
47 words
The method according to any one of claims 12 to 19, wherein said labeled antibody is labeled with an enzymatic molecule, which when treated, produces a color change to provide visual evidence of the presence on said membrane of said label and colonies of said selected bacteria.
Claim 21
Dependent Claim
23 words
The method according to claim 20, wherein said enzymatic molecule is peroxidase, alkaline phosphatase, β-galactosidase, β-glucuronidase, α-glucosidase, β-glucosidase, α-manosidase, galactose oxidase, glucoseoxidase, hexokinase.
Claim 22
Independent Claim
21 words
The method according to claims 20 or 21, wherein said membrane is treated with a chromogenic substance for said enzymatic molecule.
References Cited (Backward Citations)
No citation records indexed.
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