# US 10,865,376 B2 · Method for detecting the presence or absence of biological particles > Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt - Number: US 10,865,376 B2 - Document ID: US10865376B2 - Status: Granted - Kind: B2 - Application number: 15/319,643 - Filed: 2015-06-29 - Published: 2017-05-18 - Granted: 2020-12-15 - Earliest claimed: 2014-06-30 - Assignees: Biomerieux - Applicants: bioMérieux - Inventors: Laurent Drazek; Frédéric Pinston - Web page: https://patentgrep.com/patent/US10865376B2 ## CPC - C12M 41/36: {of biomass, e.g. colony counters or by turbidity measurements (electrooptical investigation of individual particles G01N15/14, flow cytometers G01N15/1404)} - C12M 31/00: {Means for providing, directing, scattering or concentrating light (C12M41/06 takes precedence)} - C12M 41/46: {of cellular or enzymatic activity or functionality, e.g. cell viability} - 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 1/18: Testing for antimicrobial activity of a material - C12M: APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; {APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS} - 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 A method detects the presence or absence of at least one first inhibition zone having a culture of biological particles on a culture medium in the presence of a chemical agent. The method of detection has a first step of seeding the culture medium with the biological particles, a second step in which the culture medium receives the chemical agent, a third step of incubation of the culture medium, and a fourth step of measurement of the first inhibition zone of the culture medium, which includes a first phase of taking an image of the first inhibition zone by ombroscopy. ## Claims ### Claim 1 A method for detecting the presence or absence of at least one first inhibition zone comprised by a culture of biological particles on a culture medium in the presence of a chemical agent, said method of detection comprising a first step of seeding the culture medium with the biological particles, a second step in which the culture medium receives a support impregnated with the chemical agent, and a third step of incubation of the culture medium, wherein the method of detection comprises a fourth step of measurement comprising: a first phase of taking an image of the culture medium by ombroscopy so as to generate a two-dimensional intensity image of the culture medium in which the first inhibition zone is darker than a proliferation zone of the biological particles, an analysis of the two-dimensional intensity image in which the presence of the first inhibition zone is detected, measuring a diameter of the first inhibition zone and a distance between the impregnated support and an edge of the first inhibition zone, and at least one operation of estimating a minimum inhibitory concentration based on determination of an abscissa of the referenced minimum inhibitory concentration, to determine sensitivity of the biological particles to the chemical agent; wherein the first phase, the analysis, the measuring and the at least one operation of estimating are implemented by a detecting device including a processor comprising a calculating means configured to implement the fourth step of measurement. ### Claim 2 (depends on 1) The method of detection as claimed in claim 1, wherein the method of detection comprises a fifth processing step, which comprises a second phase of taking a control photograph of the culture medium. ### Claim 3 (depends on 2) The method of detection as claimed in claim 2, wherein the control photograph is a photograph of a ruler arranged on the culture medium. ### Claim 4 (depends on 2) The method of detection as claimed in claim 2, wherein the method of detection comprises a third phase of superimposing the image and the control photograph to form a normed representation. ### Claim 5 (depends on 2) The method of detection as claimed in claim 2, wherein the fifth processing step comprises a fourth phase of localization of a reference of the ruler. ### Claim 6 (depends on 2) The method of detection as claimed in claim 2, wherein the fifth processing step comprises a fifth phase of processing the normed representation. ### Claim 7 (depends on 6) The method of detection as claimed in claim 6, wherein the fifth phase comprises at least one smoothing operation for homogenizing the normed representation. ### Claim 8 (depends on 6) The method of detection as claimed in claim 6, wherein the fifth phase comprises at least one operation of dynamic stretching of the normed representation, which comprises spreading of the normed representation to form a histogram of a contrast of the normed representation. ### Claim 9 (depends on 6) The method of detection as claimed in claim 6, wherein the fifth phase comprises at least one thresholding operation of the normed representation, which comprises detection of a threshold and determination of a contour of the normed representation. ### Claim 10 (depends on 2) The method of detection as claimed in claim 2, wherein the first phase of taking the image and the second phase of taking the control photograph are phases that are performed continuously. ### Claim 11 (depends on 2) The method of detection as claimed in claim 2, wherein the first phase of taking the image and the second phase of taking the control photograph are phases that are performed with a defined time lapse. ### Claim 12 (depends on 1) The method of detection as claimed in claim 1, wherein the first inhibition zone is present, and the presence of the first inhibition zone is detectable within four hours after the biological particles and the chemical agent are brought together. ### Claim 13 (depends on 12) The method of detection as claimed in claim 12, wherein the estimated minimum inhibitory concentration is obtained within a period of four to eight hours after the biological particles and the chemical agent are brought together.