# US 9,435,739 B2 · Methods and compositions to detect a biological activity

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- Number: US 9,435,739 B2
- Document ID: US9435739B2
- Status: Granted
- Kind: B2
- Application number: 13/883,620
- Filed: 2011-10-28
- Published: 2013-09-05
- Granted: 2016-09-06
- Earliest claimed: 2010-11-24
- Assignees: 3M Innovative Properties Company
- Applicants: Stephen B. Roscoe; Kurt J. Halverson; Jesse D. Miller; Stephanie J. Moeller; Jason W. Bjork
- Inventors: Stephen B. Roscoe; Kurt J. Halverson; Jesse D. Miller; Stephanie J. Moeller; Jason W. Bjork
- Web page: https://patentgrep.com/patent/US9435739B2

## CPC

- G01N 21/6486: {Measuring fluorescence of biological material, e.g. DNA, RNA, cells (G01N21/6428 takes precedence)}
- C12Q 1/04: Determining presence or kind of microorganism; Use of selective media for testing antibiotics or bacteriocides; Compositions containing a chemical indicator therefor {(C12Q1/6897 takes precedence)}
- G01N: INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES (measuring or testing apparatus or processes other than immunoassay, involving enzymes or microorganisms C12M, C12Q)
- C12Q: MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS (immunoassay G01N33/53); COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES

## Abstract

Compositions that comprise water, a first indicator reagent that can be converted by a first biological activity to a first biological derivative, and a plurality of particles are provided. The first indicator reagent can comprise a fluorogenic enzyme substrate having a fluorophore selected from the group consisting of umbelliferone, 7-aminocoumarin, β-naphthylamine, β-naphthol, fluorescein, resorufin, 9H-(1,3-dichloro-9,9-dimethyl acridin-2-one), rhodamine 110, a derivative of any of the foregoing fluorophores, and a combination of any of the foregoing fluorophores. The particles are capable of receiving and retaining the first biological derivative from an aqueous liquid. The first biological derivative can be indicative of a microorganism. The compositions further can comprise a gelling agent. Methods of using the compositions to detect the presence or absence of a microorganism in a sample are also provided.

## Claims

### Claim 1

A method of detecting a biological activity, comprising:
providing,
a sample that may comprise a first biological activity;
a first indicator reagent that can be converted by the biological activity to a first biological derivative, wherein the first indicator reagent comprises a fluorogenic enzyme substrate having a fluorophore selected from the group consisting of umbelliferone, 7-aminocoumarin, β-naphthylamine, β-naphthol, fluorescein, resorufin, 9H-(1,3-dichloro-9,9-dimethyl acridin-2-one), rhodamine 110, a derivative of any of the foregoing fluorophores, and a combination of any of the foregoing fluorophores;
a plurality of particles, each particle comprising an organic polymer that receives and retains the first biological derivative, wherein the organic polymer is selected from the group consisting of acrylate polymers, polystyrene polymers, cellulosic polymers, dextran polymers, a combination of any of the foregoing and a derivative of any of the foregoing; and
a gelling agent;
forming a hydrogel comprising the first indicator reagent and the gelling agent;
distributing the particles on at least a portion of a major surface of the hydrogel;
bringing the sample into fluid communication with a portion of the hydrogel, wherein the first biological derivative partitions from hydrogel onto and/or into the plurality of particles; and
detecting a presence or absence of the first biological derivative, wherein the first biological derivative is retained by a particle, wherein the presence of the first biological derivative is indicative of the presence of a first biological activity, wherein detecting the presence of the first biological derivative comprises counting a quantity of discrete fluorescent zones, and
wherein the particles are between about 0.05 microns and about 20 microns.

### Claim 2 (depends on 1)

The method of claim 1, further comprising:
providing a second indicator reagent that can be converted by a second biological activity to a second biological derivative;
bringing the second indicator into fluid communication with the hydrogel; and
detecting a presence or absence of the second biological derivative;
wherein the presence of the second biological derivative is indicative of the presence of the first biological activity.

### Claim 3 (depends on 1)

The method of claim 1, further comprising:
providing a second indicator reagent that can be converted by a second biological activity to a second biological derivative;
bringing the second indicator into fluid communication with the hydrogel; and
detecting a presence or absence of the second biological derivative.