# US 11,906,497 B2 · Rapid bacteria test

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- Number: US 11,906,497 B2
- Document ID: US11906497B2
- Status: Granted
- Kind: B2
- Application number: 17/034,800
- Filed: 2020-09-28
- Published: 2021-01-21
- Granted: 2024-02-20
- Earliest claimed: 2015-08-17
- Assignees: Swiss Analyze Bacteria AG
- Applicants: Swiss Analyze Bacteria AG
- Inventors: Dieter Ebert; Mathias Reichl; Werner Lubitz
- Web page: https://patentgrep.com/patent/US11906497B2

## CPC

- G01N 33/1826: {Organic contamination in water}
- G01N 33/537: with separation of immune complex from unbound antigen or antibody
- G01N 33/56911: {Bacteria}
- G01N 33/582: {with fluorescent label}
- G01N 21/03: Cuvette constructions
- G01N 21/65: Raman scattering
- G01N 2333/195: from bacteria
- Y02A 50/30: Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change
- 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)
- Y02A: TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE

## Abstract

The invention relates to methods and devices for quantitative determination of bacteria.

## Claims

### Claim 1

A method for quantitatively determining bacteria in a sample, comprising the steps of:
(a) incubating the sample to be tested together with a labelling reagent that can bind to the bacteria to be determined in a measuring cell comprising a first and a second chamber, wherein the labelling reagent is a labelled bacteriophage, wherein said sample is incubated in the first chamber of said measuring cell,
(b) removing labelling reagent that has not bound to the bacteria from the bacteria by means of a membrane arranged between the first and second chamber, wherein said membrane is impermeable to the bacteria to be determined and is permeable to the labelling reagent that is not bound to the bacteria to be determined, wherein the bacteria to be determined are collected on the membrane and said labelling reagent that has not bound to the bacteria is moved into said second chamber,
(c) detecting the labelling reagent bound to the bacteria collected on said membrane, and
(d) quantitatively evaluating the measurement signal from step (c), wherein the quantitative evaluation of the measurement signal comprises a calibration by using an internal standard.

### Claim 2 (depends on 1)

The method according to claim 1, wherein the bacteria are selected from the group consisting of legionella, salmonella, listeria, coliform germs, and hospital germs.

### Claim 3 (depends on 1)

The method according to claim 1, wherein said membrane has a pore diameter of approximately 0.5 to 3 μm.

### Claim 4 (depends on 1)

The method according to claim 1, wherein the labelling reagent and the sample to be tested are mixed before and/or during the incubation procedure in step (a).

### Claim 5 (depends on 1)

The method according to claim 1, wherein the labelling reagent bound to the bacteria is detected in step (c) using a laser with a confocal optic as the light source.

### Claim 6 (depends on 5)

The method according to claim 5, wherein the laser is used in an integrated device together with a spectrometer so as to form a detection unit.

### Claim 7 (depends on 1)

The method according to claim 1, wherein the quantitative evaluation in step (d) comprises sorting a specific number of bacteria into one or more categories on the basis of predetermined threshold values.

### Claim 8 (depends on 1)

The method according to claim 1, wherein at least one reference signal is used for the quantitative evaluation in step (d).

### Claim 9 (depends on 1)

The method according to claim 1, wherein the internal standard comprises fluorescent, luminescent or Raman-active particles, which bear a label that is different from said labelling regent.

### Claim 10 (depends on 1)

The method according to claim 1, wherein the labelling reagent is selected from the group consisting of a fluorescence-labelled bacteriophage, a luminescence-labelled bacteriophage and a Raman-labelled bacteriophage and wherein said labelling reagent bound to the bacteria collected, is detected by means of fluorescence measurement, luminescence measurement and/or Raman measurement.

### Claim 11 (depends on 2)

The method according to claim 2, wherein the coliform germs are Escherichia coli, and the hospital germs are Methicillin-resistant Staphylococcus aureus.

### Claim 12 (depends on 1)

The method according to claim 1, wherein said first chamber contains a store for the labelling reagent.

### Claim 13 (depends on 8)

The method according to claim 8, wherein said at least one reference signal is generated by a laser.