Method for detecting the presence or absence of biological particles

Patent metadata

Field	Value
Publication number	US-10865376-B2
Application number	US-201515319643-A
Country	US
Kind code	B2
Filing date	Jun 29, 2015
Priority date	Jun 30, 2014
Publication date	Dec 15, 2020
Grant date	Dec 15, 2020

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.

First claim

The invention claimed is:

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.

2. 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.

3. The method of detection as claimed in claim 2 , wherein the control photograph is a photograph of a ruler arranged on the culture medium.

4. 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.

5. 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.

6. The method of detection as claimed in claim 2 , wherein the fifth processing step comprises a fifth phase of processing the normed representation.

7. The method of detection as claimed in claim 6 , wherein the fifth phase comprises at least one smoothing operation for homogenizing the normed representation.

8. 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.

9. 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.

10. 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.

11. 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.

12. 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.

13. 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.

Assignees

• Biomerieux Sa

Inventors

• Drazek Laurent
• Pinston Frédéric

Classifications

• C12M41/46

  of cellular or enzymatic activity or functionality, e.g. cell viability · CPC title

• C12Q1/18

  Testing for antimicrobial activity of a material · CPC title

• C12M41/36 Primary

  of biomass, e.g. colony counters or by turbidity measurements (electrooptical investigation of individual particles G01N15/14, flow cytometers G01N15/1404) · CPC title

• C12Q1/04

  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)} · CPC title

• C12M31/00

  Means for providing, directing, scattering or concentrating light (C12M41/06 takes precedence) · CPC title