US3902969A
Expired / Lapsed
US
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Method for the identification of {i streptococcus mutans
Assignee: ADA Forsyth Institute Inc
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Filing: 1973-07-16
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Granted: 1975-09-02
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Expires: 1993-07-16
Abstract
Abstract A method for identification of Streptococcus mutans colonies growing among other oral streptococci on mitis salivarius agar has been developed, wherein colonies of Strep. mutans stain dark pink when sprayed with solutions of mannitol or sorbitol and 2,3,5 triphenyl tetrazolium chloride, while colonies of the other streptococci remain blue.
Inventors
- 👤 Olga G Gold
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-11-03 11:14
CPC Classifications
C; C12; C12Q; C12Q1/00; C12Q1/02; C12Q1/04; C12Q1/14; C12Q2304/00; C12Q2304/20; C12Q2304/24; G; G01; G01N; G01N2333/00; G01N2333/195; G01N2333/315; Y; Y10; Y10S; Y10S435/00; Y10S435/80; Y10S435/8215; Y10S435/822; Y10S435/885
Claims Analysis
Total Claims: 24
Claim 1
Independent Claim
91 words
A METHOD OF IDENTIFYING STREPTOCOCCUS MUTANS, WHICH METHOD COMPRISES: A. INOCULATING A SELECTIVE GROWTH MEDIUM FOR STRETOCOCCI WITH A BACTERIA SAMPLE CONTAINING STREPTOCOCCUS MUTANS, B. INCUBATING THE INOCULATED MEDIUM TO PROVIDE STREP.MUTANS COLONIES, C. CONTACTING THE STREP.MUTANS WITH MANNITOL OR SORBITOL, OR COMBINATIONS THEREOF, AND FERMENTING THE MANNITOL OR SORBITOL IN THE PRESENCE OF AN OXIDATION-REDUCTION INDICATOR WHICH CHANGES COLOR ON THE FERMENTATION OF THE MANNITOL OR SORBITOL BY THE STREP.MUTANS, AND D. IDENTIFYING THE STREP.MUTANS BY THE DIFFERENTIAL STAINING OF THE STREP.MUTANS COLONIES BY CHANGE IN COLOR OF THE INDICATOR.
Claim 2
Independent Claim
91 words
A METHOD OF IDENTIFYING STREPTOCOCCUS MUTANS, WHICH METHOD COMPRISES: A. INOCULATING A SELECTIVE GROWTH MEDIUM FOR STRETOCOCCI WITH A BACTERIA SAMPLE CONTAINING STREPTOCOCCUS MUTANS, B. INCUBATING THE INOCULATED MEDIUM TO PROVIDE STREP.MUTANS COLONIES, C. CONTACTING THE STREP.MUTANS WITH MANNITOL OR SORBITOL, OR COMBINATIONS THEREOF, AND FERMENTING THE MANNITOL OR SORBITOL IN THE PRESENCE OF AN OXIDATION-REDUCTION INDICATOR WHICH CHANGES COLOR ON THE FERMENTATION OF THE MANNITOL OR SORBITOL BY THE STREP.MUTANS, AND D. IDENTIFYING THE STREP.MUTANS BY THE DIFFERENTIAL STAINING OF THE STREP.MUTANS COLONIES BY CHANGE IN COLOR OF THE INDICATOR.
Claim 3
Dependent Claim
14 words
The method of claim 1 wherein the selective growth medium is mitis salivarius agar.
Claim 4
Dependent Claim
19 words
The method of claim 1 wherein the bacteria sample is a sample from the oral cavity of a patient.
Claim 5
Dependent Claim
12 words
The method of claim 1 wherein the indicator is triphenyl tetrazolium chloride.
Claim 6
Dependent Claim
21 words
The method of claim 1 wherein the mannitol or sorbitol and the indicator are sprayed onto the medium containing the Strep.mutans.
Claim 7
Dependent Claim
28 words
The method of claim 5 wherein the medium is first sprayed with an aqueous mannitol or sorbitol-containing solution and, after incubation, sprayed with a triphenyl tetrazolium chloride solution.
Claim 8
Dependent Claim
24 words
The method of claim 1 wherein the mannitol or sorbitol and the indicator are separately applied as solutions and aerobically incubated after each application.
Claim 9
Dependent Claim
17 words
The method of claim 1 wherein incubating the medium is carried out, first, anaerobically, and then aerobically.
Claim 10
Dependent Claim
17 words
The method of claim 1 wherein the inoculated incubated stained colonies consist essentially of Strep.mutans and enterococci.
Claim 11
Dependent Claim
31 words
The method of claim 1 wherein the mannitol or sorbitol and the indicator are applied to the Strep.mutans colonies in an aqueous phosphate buffered solution having a pH of about 7.4.
Claim 12
Independent Claim
66 words
A method of identifying Streptococcus mutans. which method comprises: a. contacting Strep.mutans colonies on a mitis salivarius agar growth medium with mannitol or sorbitol, or combinations thereof, and a triphenyl tetrazolium chloride indicator and fermenting the mannitol or sorbitol by the Strep.mutans and to obtain a change in color of the indicator from colorless to red; and b. identifying the colonies of Strep.mutans on the agar.
Claim 13
Independent Claim
66 words
A method of identifying Streptococcus mutans. which method comprises: a. contacting Strep.mutans colonies on a mitis salivarius agar growth medium with mannitol or sorbitol, or combinations thereof, and a triphenyl tetrazolium chloride indicator and fermenting the mannitol or sorbitol by the Strep.mutans and to obtain a change in color of the indicator from colorless to red; and b. identifying the colonies of Strep.mutans on the agar.
Claim 14
Dependent Claim
43 words
The method of claim 11 wherein the growth medium is inoculated with a bacteria sample from the oral cavity of a patient; and wherein the step of identifying includes determining the number of Strep.mutans to the total oral streptococcal flora of the sample.
Claim 15
Dependent Claim
32 words
The method of claim 11 wherein the mannitol or sorbitol and the indicator are separately sprayed in aqueous solutions onto the surface of the growth medium, and incubated after each such spraying.
Claim 16
Dependent Claim
27 words
The method of claim 11 wherein the mannitol or sorbitol and the triphenyl tetrazolium chloride are in phosphate buffered aqueous solutions having a pH of about 7.4.
Claim 17
Dependent Claim
37 words
An aqueous solution useful for the identification of Streptococcus mutans which comprises in combination: a. mannitol or sorbitol or a combination thereof; and b. triphenyl tetrazolium chloride, the solution buffered at a pH of greater than 6.0.
Claim 18
Dependent Claim
37 words
An aqueous solution useful for the identification of Streptococcus mutans which comprises in combination: a. mannitol or sorbitol or a combination thereof; and b. triphenyl tetrazolium chloride, the solution buffered at a pH of greater than 6.0.
Claim 19
Dependent Claim
17 words
The solution of claim 15 which is a phosphate buffered solution having a pH of about 7.4.
Claim 20
Dependent Claim
38 words
The solution of claim 15 wherein the mannitol or sorbitol is present in an amount of up to about 0.1 to 20% by weight, and the triphenyl tetrazolium chloride in an amount of 0.1 to 5.0% by weight.
Claim 21
Dependent Claim
44 words
An aqueous solution useful for the identification of Streptococcus mutans which comprises in combination: up to about 10% by weight of mannitol or sorbitol, or combinations thereof, and triphenyl tetrazolium chloride as an indicator, the solution phosphate buffered at a pH of about 7.4.
Claim 22
Dependent Claim
44 words
An aqueous solution useful for the identification of Streptococcus mutans which comprises in combination: up to about 10% by weight of mannitol or sorbitol, or combinations thereof, and triphenyl tetrazolium chloride as an indicator, the solution phosphate buffered at a pH of about 7.4.
Claim 23
Independent Claim
122 words
A method of identifying Strep.mutans and differentiating Strep.mutans from other streptococci and enterococci, which method comprises: a. providing a medium containing thereon colonies of Strep.mutans and other organisms, the colonies derived from a sample taken froM the oral cavity of a patient; b. contacting the colonies on the medium with an aqueous solution comprising mannitol or sorbitol or a combination thereof and triphenyl tetrazolium chloride in the colorless reduced state; c. incubating the colonies so contacted, and fermenting selectively the mannitol or sorbitol by the Strep.mutans to oxidize and change the color of the triphenyl tetrazolium chloride, thereby staining the Strep.mutans colonies a dark pink color; and d. identifying and differentiating the stained Strep.mutans colonies from the other organisms on the medium.
Claim 24
Independent Claim
122 words
A method of identifying Strep.mutans and differentiating Strep.mutans from other streptococci and enterococci, which method comprises: a. providing a medium containing thereon colonies of Strep.mutans and other organisms, the colonies derived from a sample taken froM the oral cavity of a patient; b. contacting the colonies on the medium with an aqueous solution comprising mannitol or sorbitol or a combination thereof and triphenyl tetrazolium chloride in the colorless reduced state; c. incubating the colonies so contacted, and fermenting selectively the mannitol or sorbitol by the Strep.mutans to oxidize and change the color of the triphenyl tetrazolium chloride, thereby staining the Strep.mutans colonies a dark pink color; and d. identifying and differentiating the stained Strep.mutans colonies from the other organisms on the medium.
References Cited (Backward Citations)
USPTO Annuity & Maintenance Windows
| Annuity Stage | Window Opens | Payment Deadline | Surcharge Period Until | Estimated Fee | Status |
|---|---|---|---|---|---|
| 3.5 Years (Year 4) | 1978-09-01 | 1979-03-03 | 1979-09-02 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 1982-09-01 | 1983-03-03 | 1983-09-02 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 1986-09-01 | 1987-03-03 | 1987-09-02 | $7700.00 | Window Closed / Lapsed |