# US 11,898,192 B2 · Method for quantifying at least one microorganism group via mass spectrometry

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- Number: US 11,898,192 B2
- Document ID: US11898192B2
- Status: Granted
- Kind: B2
- Application number: 15/503,802
- Filed: 2015-08-11
- Published: 2017-09-28
- Granted: 2024-02-13
- Earliest claimed: 2014-08-14
- Assignees: Biomerieux
- Applicants: BioméRieux
- Inventors: Chloé Bardet; Tiphaine Cecchini; Yannick Charretier; Jean-Philippe Charrier; Christelle Compagnon; Tanguy Fortin
- Web page: https://patentgrep.com/patent/US11898192B2

## CPC

- C12Q 1/06: Quantitative determination
- G01N 33/569: for microorganisms, e.g. protozoa, bacteria, viruses
- G01N 33/6848: {Methods of protein analysis involving mass spectrometry}
- 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
- 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)

## Abstract

A method for quantifying at least one microorganism group via at least one mass spectrometry analysis. The method includes at least one separation and fragmentation step. The method moreover includes a step that involves measuring the amount of at least one representative peptide or at least one protein representing the microorganism group. The at least one representative peptide or the at least one protein is obtained after the at least one separation and fragmentation step and serves as a quantification marker(s). The amount of the quantification marker(s) is directly correlatable to the amount of the at least one microorganism group.

## Claims

### Claim 1

A method for quantifying a microorganism from Escherichia coli species in a biological sample by mass spectrometry, the method comprising:
treating proteins of the microorganism by alkylation and digestion in order to fragment the proteins, thereby obtaining peptides including a proteotypic peptide having the amino acid sequence of SEQ ID NO:2, wherein the proteotypic peptide is representative of the Escherichia coli species and does not vary in amount due to the growth phase of the microorganism;
measuring an amount of the proteotypic peptide, as a quantification marker, using a mass spectrometer comprising a detector; and
quantifying the microorganism based on an electrical signal produced by the detector.

### Claim 2 (depends on 1)

The method of claim 1, wherein the electrical signal is processed using a computer to quantify the microorganism.

### Claim 3 (depends on 1)

The method of claim 1, wherein the mass spectrometry is PRM, SRM, MRM, MS2, MRM3, DDA (data dependent acquisition) or DIA (data independent acquisition) mass spectrometry.

### Claim 4 (depends on 1)

The method of claim 1, performed in less than one hour.