Methods and articles for detecting deoxyribonuclease activity

Patent metadata

Field	Value
Publication number	US-9738919-B2
Application number	US-201414529481-A
Country	US
Kind code	B2
Filing date	Oct 31, 2014
Priority date	Feb 26, 2009
Publication date	Aug 22, 2017
Grant date	Aug 22, 2017

Abstract

The disclosure provides articles and methods useful for detecting a discrete source of DNase activity. DNase-producing microorganisms can be detected. The device can further include selective agents and/or indicators to differentiate groups or species microorganisms. Methods of use include detecting or enumerating DNase-producing microorganisms.

First claim

What is claimed is:

1. A method of detecting deoxyribonuclease activity, comprising:
providing
a hydrogel comprising a DNase indicator system including methyl green and DNA, wherein the concentration of methyl green in the hydrogel is at least about 5 μg/mL to about 20 μg/mL;
a sample suspected of containing a discrete source of deoxyribonuclease activity; and
an imaging system;

contacting, for a predetermined period of time, i) a predetermined volume of aqueous liquid comprising the sample and ii) the hydrogel;
wherein after said contacting the predetermined volume of the aqueous liquid and the hydrogel, the concentration of said methyl green in the hydrogel is about 5 ug/mL to 20 μg/mL;
obtaining an image of the hydrogel with the imaging system; and
detecting a discrete source of deoxyribonuclease activity, wherein said detecting a discrete source of deoxyribonuclease activity comprises displaying, printing, or analyzing the image of the hydrogel.

2. The method of claim 1 , wherein the discrete source of deoxyribonuclease activity is a microorganism.

3. The method of claim 1 , wherein the hydrogel further comprises a nutrient, a selective agent, an antibiotic, or an indicator.

4. The method of claim 1 , further comprising the step of providing a nutrient, a selective agent, an indicator, or any combination of two or more of the foregoing; wherein said contacting for a predetermined period of time comprises contacting the sample of step i) and the hydrogel of step ii) with the nutrient, the selective agent, the indicator, or any combination of two or more of the foregoing.

5. The method of claim 1 , wherein the imaging system comprises an illumination source and wherein obtaining an image of the hydrogel comprises illuminating the hydrogel.

6. The method of claim 5 , wherein illuminating the hydrogel comprises illuminating the hydrogel with a limited band of visible wavelengths.

7. The method of claim 5 , herein illuminating the hydrogel comprises using a bias filter to illuminate the hydrogel or to collect the image of the hydrogel.

8. The method of claim 1 , further comprising the step of providing an image analysis system; wherein said analyzing the image comprises analyzing the image with the image analysis system.

9. The method of claim 8 , wherein said analyzing the image comprises analyzing selected wavelengths of the image.

Assignees

• 3M Innovative Properties Co

Inventors

• Mach Patrick A
• Rosauer Michelle L
• Hughes Michael E

Classifications

• C12Q1/34 Primary

  involving hydrolase · CPC title

• G01N2333/31

  from Staphylococcus (G) · CPC title

• C12Q1/04 Primary

  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)} · CPC title

• C12N11/04

  entrapped within the carrier, e.g. gel or hollow fibres · CPC title

• G01N2333/922

  Ribonucleases (RNAses); Deoxyribonucleases (DNAses) · CPC title