US4126516A
Expired / Lapsed
US
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Method for distinguishing gram positive and gram negative microbes
Assignee: Corning Glass Works
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Filing: 1976-11-29
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Granted: 1978-11-21
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Expires: 1996-11-29
Abstract
Abstract Microbial culture device containing a growth medium having incorporated therein a lipophilic fluorescent material and means associated therewith for localizing and identifying the growth of colonies of an unknown microbe, and a known gram positive microbe, and/or a known gram negative microbe. The device can be used to determine whether an unknown microbe sample is gram positive or gram negative by culturing the unknown on the medium and then comparing the fluorescence of the unknown microbes with fluorescence of similarly cultured gram positive and/or gram negative microbes.
Inventors
- 👤 Ralph A. Messing
- 👤 William S. Ramsey
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-10-30 21:27
CPC Classifications
C; C12; C12Q; C12Q1/00; C12Q1/02; C12Q1/04; Y; Y10; Y10S; Y10S435/00; Y10S435/8215; Y10S435/822; Y10S435/83; Y10S435/843; Y10S435/848; Y10S435/849; Y10S435/852; Y10S435/873; Y10S435/874; Y10S435/875; Y10S435/88
Claims Analysis
Total Claims: 24
Claim 1
Independent Claim
90 words
A method of determining the gram staining properties of a microbe, the method comprising the steps of: (a) incubating a sample of the microbe on a microbial culture medium having incorporated therein a lipophilic fluorescent material in an amount sufficient to permit the fluorometric detection of microbial growth; (b) determining the quantity of fluorescence of the incubated microbe; and (c) comparing the quantity of fluorescence of the incubated microbial sample with the quantity of fluorescence of similarly incubated microbe known to be a gram positive or a gram negative microbe.
Claim 2
Independent Claim
90 words
A method of determining the gram staining properties of a microbe, the method comprising the steps of: (a) incubating a sample of the microbe on a microbial culture medium having incorporated therein a lipophilic fluorescent material in an amount sufficient to permit the fluorometric detection of microbial growth; (b) determining the quantity of fluorescence of the incubated microbe; and (c) comparing the quantity of fluorescence of the incubated microbial sample with the quantity of fluorescence of similarly incubated microbe known to be a gram positive or a gram negative microbe.
Claim 3
Dependent Claim
18 words
The method of claim 1 wherein the microbial culture medium used in step (a) comprises a gel-like material.
Claim 4
Dependent Claim
11 words
The method of claim 2 wherein the medium comprises an agar.
Claim 5
Dependent Claim
18 words
The method of claim 1 wherein the lipophilic fluorescent material comprises 8-anilino-1-naphthalene sulfonic acid or a salt thereof.
Claim 6
Dependent Claim
18 words
The method of claim 1 wherein the lipophilic fluorescent material comprises 8-p-toluidino-1-naphthalene sulfonic acid or a salt thereof.
Claim 7
Dependent Claim
18 words
The method of claim 1 wherein the lipophilic fluorescent material comprises 6-p-toluidino-1-naphthalene sulfonic acid or a salt thereof.
Claim 8
Dependent Claim
13 words
The method of claim 1 wherein the lipophilic fluorescent material comprises acridine orange.
Claim 9
Dependent Claim
17 words
The method of claim 1 wherein the lipophilic fluorescent material comprises 1-pyrenebutyric acid or a salt thereof.
Claim 10
Dependent Claim
33 words
The method of claim 1 wherein the microbial culture medium comprises an agar and the lipophilic fluorescent material is 8-anilino-1-naphthalene-sulfonic acid in an amount ranging from about 0.08 to 0.5 mg/ml of agar.
Claim 11
Dependent Claim
22 words
The method of claim 1 wherein the fluorescence quantity of step (b) is determined using an aqueous suspension of the incubated sample.
Claim 12
Independent Claim
105 words
A method of determining whether the lipid content of the cell wall of a given microbe is above or below the lipid content of the cell wall of a microbe used as a reference, the method comprising the steps of: (a) incubating the given microbe on a microbial growth medium having incorporated therein a lipophilic fluorescent material in an amount sufficient to permit the fluorometric detection of microbial growth; (b) determining the quantity of fluorescence of the incubated microbe; and (c) comparing the quantity of fluorescence of step (b) with the quantity of fluorescence emitted by the reference microbe when incubated according to step (a).
Claim 13
Independent Claim
105 words
A method of determining whether the lipid content of the cell wall of a given microbe is above or below the lipid content of the cell wall of a microbe used as a reference, the method comprising the steps of: (a) incubating the given microbe on a microbial growth medium having incorporated therein a lipophilic fluorescent material in an amount sufficient to permit the fluorometric detection of microbial growth; (b) determining the quantity of fluorescence of the incubated microbe; and (c) comparing the quantity of fluorescence of step (b) with the quantity of fluorescence emitted by the reference microbe when incubated according to step (a).
Claim 14
Dependent Claim
14 words
The method of claim 11 wherein the reference microbe is a gram negative microbe.
Claim 15
Dependent Claim
14 words
The method of claim 11 wherein the reference microbe is a gram positive microbe.
Claim 16
Dependent Claim
13 words
The method of claim 11 wherein the microbial growth medium comprises a fluid.
Claim 17
Dependent Claim
13 words
The method of claim 11 wherein the microbial growth medium comprises gel-like material.
Claim 18
Dependent Claim
12 words
The method of claim 15 wherein the growth medium comprises an agar.
Claim 19
Dependent Claim
18 words
The method of claim 11 wherein the lipophilic fluorescent material comprises 8-anilino-1-naphthalene sulfonic acid or a salt thereof.
Claim 20
Dependent Claim
18 words
The method of claim 11 wherein the lipophilic fluorescent material comprises 8-p-toluidino-1-naphthalene sulfonic acid or a salt thereof.
Claim 21
Dependent Claim
18 words
The method of claim 11 wherein the lipophilic fluorescent material comprises 6-p-toluidino-2-naphthalene sulfonic acid or a salt thereof.
Claim 22
Dependent Claim
13 words
The method of claim 11 wherein the lipophilic fluorescent material comprises acridine orange.
Claim 23
Dependent Claim
17 words
The method of claim 11 wherein the lipophilic fluorescent material comprises 1-pyrenebutyric acid or a salt thereof.
Claim 24
Dependent Claim
39 words
The method of claim 11 wherein the microbial growth medium comprises agar and the lipophilic fluorescent material is 8-anilino-1-naphthalene sulfonic acid or a salt thereof in an amount ranging from about 0.08 to 0.5 mg per ml of agar.
References Cited (Backward Citations)
US3165450A
Cited
USPTO Annuity & Maintenance Windows
| Annuity Stage | Window Opens | Payment Deadline | Surcharge Period Until | Estimated Fee | Status |
|---|---|---|---|---|---|
| 3.5 Years (Year 4) | 1981-11-20 | 1982-05-22 | 1982-11-21 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 1985-11-20 | 1986-05-22 | 1986-11-21 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 1989-11-20 | 1990-05-22 | 1990-11-21 | $7700.00 | Window Closed / Lapsed |