# US 8,785,142 B2 · Test arrangement > Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt - Number: US 8,785,142 B2 - Document ID: US8785142B2 - Status: Granted - Kind: B2 - Application number: 13/577,366 - Filed: 2011-02-09 - Published: 2012-12-06 - Granted: 2014-07-22 - Earliest claimed: 2010-02-10 - Assignees: Eva Sigl - Applicants: Georg Gübitz; Eva Sigl; Andrea Hasmann; Marc Schröder; Konstantin Schneider; Alexandra Rollett; Franz Kaufmann; Andreas Hafner - Inventors: Georg Gübitz; Eva Sigl; Andrea Hasmann; Marc Schröder; Konstantin Schneider; Alexandra Rollett; Franz Kaufmann; Andreas Hafner - Web page: https://patentgrep.com/patent/US8785142B2 ## CPC - C12Q 1/001: {Enzyme electrodes} - 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)} - 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 The invention relates to an arrangement, comprising a solid carrier and a matrix arranged on the solid carrier, said matrix comprising at least one enzymatically convertible or modifiable molecule and comprising at least one enzyme that can be released by the conversion or modification of the molecule, said enzyme being capable of converting at least one color-changing substrate located in the matrix and/or on the solid carrier. ## Claims ### Claim 1 An arrangement comprising a solid carrier and a matrix arranged on the solid carrier, the matrix comprising at least one trigger enzyme convertible or modifiable molecule and at least one amplifier enzyme that can be released by conversion or modification of the molecule, wherein the amplifier enzyme converts at least one colour-changing immobilized substrate located in the matrix and/or on the solid carrier after the amplifier enzyme is released. ### Claim 2 (depends on 1) The arrangement according to claim 1, characterised in that the matrix is arranged as a layer, in the form of capsules or as a hydrogel on the solid carrier. ### Claim 3 (depends on 1) The arrangement according to claim 1, characterised in that the enzymatically convertible or modifiable molecule is a polymer. ### Claim 4 (depends on 3) The arrangement according to claim 3, characterised in that the polymer is a polysaccharide, polypeptide, polyester, polyamide or a combination thereof. ### Claim 5 (depends on 4) The arrangement according to claim 4, characterised in that the polysaccharide is selected from the group consisting of pectin, amylose, amylopectin, agarose, alginate, carrageenan, chitin, chitosan, dextran, glycogen, locust bean gum, levan, pollulan, xanthan and xylan. ### Claim 6 (depends on 3) The arrangement according to claim 3, characterised in that the polymer is selected from the group consisting of, agar, agarose, maltodextrins, alginic acid and salts thereof, sodium alginate, calcium alginate, collagen, chitosan, peptidoglycan, gelatins, albumin, shellac, polysaccharides, starch, dextran, cyclodextrins, pectin, and carragenan. ### Claim 7 (depends on 3) The arrangement according to claim 3, characterised in that the polymer, the amplifier enzyme and the colour-changing substrate are selected from Table A. ### Claim 8 (depends on 1) The arrangement according to claim 1, characterised in that the amplifier enzyme is conjugated to polyvinyl alcohol, polyethylene glycol (PEG), polypeptide, elastin or a peptide. ### Claim 9 (depends on 8) The arrangement according to claim 8, characterised in that the amplifier enzyme is conjugated by means of a polypeptide or peptide to the enzymatically convertible or modifiable molecule, and wherein the trigger enzyme is selected from the group consisting of a microbial enzyme, an enzyme of the immune system and elastase. ### Claim 10 (depends on 1) The arrangement according to claim 1, characterised in that the colour-changing substrate is selected from the group consisting of phenolic compounds and azo dyes. ### Claim 11 (depends on 10) The arrangement according to claim 10, characterised in that the colour-changing substrate can be converted directly by an amplifier enzymes selected from the group consisting of microbial enzymes, laccases, peroxidases, enzymes of the immune system, and myeloperoxidase. ### Claim 12 (depends on 1) The arrangement according to claim 1, characterised in that the arrangement comprises a semi-permeable membrane opposite the solid carrier. ### Claim 13 (depends on 12) The arrangement according to claim 12, characterised in that the semi-permeable membrane is selected from the group consisting of cellulose derivatives, polyamides, polyacrylamides and polyester. ### Claim 14 (depends on 1) The arrangement according to claim 1, characterised in that the solid carrier comprises a material which is selected from the group consisting of inorganic materials, silica gel, aluminium, silicon, glass, organic materials, polyester, polystyrene, polyamide, polyacrylamide, polyvinyl alcohol, biopolymers, and paper. ### Claim 15 (depends on 1) A method of determining the presence of cells of interest in a sample comprising: applying the sample to the arrangement of claim 1, and detecting the presence or absence of a change in colour in the colour-changing substrate, wherein the convertible or modifiable molecule of the arrangement is modified or converted by a trigger enzyme produced by the cells of interest and the presence of a change in colour of the colour-changing substrate indicates that the cells of interest are present in the sample, and the absence of a change in colour of the colour-changing substrate indicates the cells of interest are absent in the sample. ### Claim 16 (depends on 15) The method of claim 15 wherein the cells are microorganisms. ### Claim 17 (depends on 16) The method of claim 16 wherein the microorganisms are selected from the group consisting of bacteria and fungi. ### Claim 18 (depends on 1) A method of detecting at least one trigger enzyme in a sample comprising: applying the sample to the arrangement of claim 1, and detecting the presence or absence of a change in colour of the colour-changing substrate, wherein the convertible or modifiable molecule of the arrangement is modified or converted by the at least one trigger enzyme of interest and the presence of a change in colour of the colour-changing substrate indicates the at least one trigger enzyme of interest is present in the sample, and the absence of a change in colour of the colour-changing substrate indicates the at least one trigger enzyme of interest is absent in the sample. ### Claim 19 (depends on 1) A method of detecting wound infection by determining the presence of at least one wound-specific trigger enzyme in a sample comprising: applying the sample to the arrangement of claim 1, and detecting the presence or absence of the colour-changing substrate, wherein the convertible or modifiable molecule of the arrangement is modified or converted by the at least one wound-specific trigger enzyme and the presence of a change in colour of the colour-changing substrate indicates the at least one wound-specific trigger enzyme is present in the sample, and the absence of a change in colour of the colour-changing substrate indicates the at least one wound-specific trigger enzyme is absent in the sample. ### Claim 20 (depends on 19) The method of claim 19, wherein the at least one wound-specific trigger enzyme is selected from the group consisting of amylase, cellulase, xylanase, mannanase, protease, lysozyme, elastase, collagenase, and cathepsin. ### Claim 21 (depends on 1) The arrangement according to claim 1, wherein the amplifier enzyme is selected from the group consisting of a protease, a laccase, and a peroxidase.