US4591554A
Expired / Lapsed
US
⏱️ 0.0 years remaining
Rapid method for detecting microorganisms
Assignee: Ajinomoto Co Inc
•
Filing: 1983-09-16
•
Granted: 1986-05-27
•
Expires: 2003-09-16
Abstract
Abstract A small number of microorganisms can be rapidly detected or determined by means of a fluorescence analysis method using specific umbelliferone derivatives such as 4-methylumbelliferyl phosphate and 4-methyl umbelliferyl galactoside, and this detecting method is applicable for a microbial inspection on sanitary quality of various kind of food, beverage, water and toilet article, or for a clinical inspection on a microbial infection.
Inventors
- 👤 Ichiro Koumura
- 👤 Hideo Kanou
- 👤 Masahiko Okunishi
- 👤 Kazuhiko Yamada
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-11-02 23:01
CPC Classifications
C; C12; C12Q; C12Q1/00; C12Q1/02; C12Q1/04; C12Q1/10; C12Q1/06; C12Q1/34; C12Q2334/00; C12Q2334/20; C12Q2334/22
Claims Analysis
Total Claims: 13
Claim 1
Independent Claim
131 words
A method for rapidly determining a small number of microorganisms, less than 10 4 per 1 milliliter, in a sample, which comprises: (A) forming a nutrient-containing solution consisting essentially of a nutrient medium capable of supporting propagation of the microorganisms and an aqueous solvent; (B) mixing said solution with a non-fluorescent umbelliferone derivative; (C) introducing said sample which contains microorganisms into said solution; (D) allowing said microorganisms to propagate in said solution consisting essentially of said nutrient-containing solution, umbelliferone derivative and sample for 1 to 12 hours; (E) measuring the amount of fluorescent umbelliferone derivative liberated by action of propagated microorganisms with a fluorescence detector; and (F) determining the number of microorganisms less than 10 4 per 1 milliliter of said sample based on the amount of liberated fluorescent umbelliferone derivative.
Claim 2
Independent Claim
131 words
A method for rapidly determining a small number of microorganisms, less than 10 4 per 1 milliliter, in a sample, which comprises: (A) forming a nutrient-containing solution consisting essentially of a nutrient medium capable of supporting propagation of the microorganisms and an aqueous solvent; (B) mixing said solution with a non-fluorescent umbelliferone derivative; (C) introducing said sample which contains microorganisms into said solution; (D) allowing said microorganisms to propagate in said solution consisting essentially of said nutrient-containing solution, umbelliferone derivative and sample for 1 to 12 hours; (E) measuring the amount of fluorescent umbelliferone derivative liberated by action of propagated microorganisms with a fluorescence detector; and (F) determining the number of microorganisms less than 10 4 per 1 milliliter of said sample based on the amount of liberated fluorescent umbelliferone derivative.
Claim 3
Independent Claim
148 words
A method for rapidly determining a small number of microorganisms, less than 10 4 per 1 milliliter, in a sample, which comprises: (A) forming a nutrient-containing solution consisting essentially of a nutrient medium capable of supporting propagation of the microorganisms and an aqueous solvent; (B) introducing said sample which contains microorganisms into said solution; (C) allowing said microorganism to propagate in said solution for 1 to 12 hours; (D) mixing said solution with a non-fluorescent umbelliferone derivative to provide a mixture; (E) incubating said mixture consisting essentially of said nutrient-containing solution, said sample and said non-fluorescent umbelliferone derivative for from 1 to 2 hours; (F) measuring the amount of fluorescent umbelliferone derivative liberated by action of propagated microorganisms with a fluorescence detector; and (G) determining the number of microorganisms less than 10 4 per 1 milliliter of said sample based on the amount of liberated fluorescent umbelliferone derivative.
Claim 4
Independent Claim
148 words
A method for rapidly determining a small number of microorganisms, less than 10 4 per 1 milliliter, in a sample, which comprises: (A) forming a nutrient-containing solution consisting essentially of a nutrient medium capable of supporting propagation of the microorganisms and an aqueous solvent; (B) introducing said sample which contains microorganisms into said solution; (C) allowing said microorganism to propagate in said solution for 1 to 12 hours; (D) mixing said solution with a non-fluorescent umbelliferone derivative to provide a mixture; (E) incubating said mixture consisting essentially of said nutrient-containing solution, said sample and said non-fluorescent umbelliferone derivative for from 1 to 2 hours; (F) measuring the amount of fluorescent umbelliferone derivative liberated by action of propagated microorganisms with a fluorescence detector; and (G) determining the number of microorganisms less than 10 4 per 1 milliliter of said sample based on the amount of liberated fluorescent umbelliferone derivative.
Claim 5
Dependent Claim
22 words
The method as set forth in claim 2, wherein the cells of the propagated microorganism are ruptured prior to the mixing step.
Claim 6
Independent Claim
42 words
The method as set forth in claims 1 or 2, wherein the non-fluorescent umbelliferone derivative is a compound of the formula ##STR2## wherein R 1 represents a hexose residue, phosphate, pyrosphosphate or acyl group, and R 2 represents hydrogen or methyl group.
Claim 7
Dependent Claim
30 words
The method as set forth in claim 4, wherein R 1 is glucoside residue, arabinoside residue, galactoside residue, phosphate, pyrosphosphate or acetyl group and R 2 is a methyl group.
Claim 8
Dependent Claim
20 words
The method as set forth in claim 5, wherein R 1 is phosphate and R 2 is a methyl group.
Claim 9
Independent Claim
19 words
The method as set forth in claims 1 or 2, wherein the microorganisms to be determined are coliform bacteria.
Claim 10
Dependent Claim
22 words
The method as set forth in claim 7, wherein 4-methyl umbelliferyl galactoside and/or 4-methyl umbelliferyl arabinoside is used as the umbelliferone derivative.
Claim 11
Dependent Claim
18 words
The method as set forth in claim 8, wherein an inducer of β-galactosidase is added to the sample.
Claim 12
Independent Claim
21 words
The method as set forth in claims 1 or 2, wherein a bile acid or desoxycholate is added to the sample.
Claim 13
Independent Claim
25 words
The method as set forth in claims 1 or 2, wherein the number of microorganisms is determined according to the conventional Most Probable Number method.
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) | 1989-05-26 | 1989-11-25 | 1990-05-27 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 1993-05-26 | 1993-11-25 | 1994-05-27 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 1997-05-26 | 1997-11-25 | 1998-05-27 | $7700.00 | Window Closed / Lapsed |