# US 10,480,020 B2 · Rapid test for microbial resistances by mass spectrometry

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- Number: US 10,480,020 B2
- Document ID: US10480020B2
- Status: Granted
- Kind: B2
- Application number: 15/089,653
- Filed: 2016-04-04
- Published: 2016-10-13
- Granted: 2019-11-19
- Earliest claimed: 2015-04-13
- Applicants: Bruker Daltonik GmbH
- Inventors: Katrin Sparbier; Beatrix Wegemann
- Web page: https://patentgrep.com/patent/US10480020B2

## CPC

- C12Q 1/18: Testing for antimicrobial activity of a material
- H01J 49/0418: {for laser desorption, e.g. matrix-assisted laser desorption/ionisation [MALDI] plates or surface enhanced laser desorption/ionisation [SELDI] plates}
- 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
- H01J: ELECTRIC DISCHARGE TUBES OR DISCHARGE LAMPS (spark-gaps H01T; electric arc lamps H05B31/00; particle accelerators H05H)

## Abstract

The invention relates to methods and instruments for determining the resistances of microbes to antibiotics, in particular those microbes which produce beta-lactamases. The method determines the resistance of the microbes in less than an hour by incubating a tiny quantity of the microbes on a mass spectrometric sample support plate after they have been combined with a dosed quantity of a suitable antibiotic, for example the beta-lactam antibiotic imipenem, and by direct mass spectrometric measurement of the breakdown of the antibiotic by the microbial enzymes.

## Claims

### Claim 1

Sample support for a mass spectrometric determination of the resistance of a microbial sample to be applied to the sample support, wherein the sample support plate is flat and has on its surface a multiplicity of test sites and at least one test site is coated with a dosed quantity of one or more antibiotics and with one or more reference substances.

### Claim 2 (depends on 1)

The sample support according to claim 1, wherein different test sites are coated with different antibiotics.

### Claim 3 (depends on 1)

The sample support according to claim 1, wherein the dosed quantity is around 0.1 to 2 micrograms.

### Claim 4 (depends on 1)

The sample support according to claim 1, wherein the one or more antibiotics is a beta-lactam antibiotic.

### Claim 5 (depends on 4)

The sample support according to claim 4, wherein the beta-lactam antibiotic is from the groups of penicillins, the cephalosporins, the monobactams or the carbapenems.

### Claim 6 (depends on 5)

The sample support according to claim 5, wherein the beta-lactam antibiotic is from the groups of benzylpenicillins, oral penicillins, aminopenicillins, isoxacylpenicillins, or acylaminopenicillins.

### Claim 7 (depends on 4)

The sample support according to claim 4, wherein the one or more antibiotics is a beta-lactam antibiotic in combination with a beta-lactamase inhibitor.

### Claim 8 (depends on 7)

The sample support according to claim 7, wherein the beta-lactamase inhibitor is sulbactam.

### Claim 9 (depends on 7)

The sample support according to claim 7, wherein the one or more antibiotics is amoxicillin in combination with clavulanic acid, piperacillin in combination with tazobactam or ampicillin in combination with sulbactam.

### Claim 10 (depends on 1)

The sample support according to claim 1, wherein said mass spectrometric determination is a mass spectrometric determination using matrix assisted laser desorption/ionization (MALDI).

### Claim 11 (depends on 1)

The sample support according to claim 1, wherein the tests sites have a capacity of less than 10 microliters.

### Claim 12 (depends on 11)

The sample support according to claim 11, wherein the tests sites have a capacity between one and three microliters.

### Claim 13 (depends on 1)

The sample support according to claim 1, wherein the number of test sites is between 48 and 386.

### Claim 14 (depends on 1)

The sample support according to claim 1, wherein the test sites are hydrophilic with hydrophobic surroundings.