Enrichment and selective culture of mycobacteria

Patent metadata

Field	Value
Publication number	US-10597693-B2
Application number	US-201615549370-A
Country	US
Kind code	B2
Filing date	Feb 4, 2016
Priority date	Feb 6, 2015
Publication date	Mar 24, 2020
Grant date	Mar 24, 2020

Abstract

A process for the enrichment and selective culture of mycobacteria contained in a biological sample, wherein all or part of the sample is inoculated in/on a culture medium including a nutritive component suitable for the development and growth of mycobacteria, wherein the culture medium includes, as selective agents, at least 9-chloro-9-(4′-diethylamino)phenyl-9,10-dihydro-10-phenylacridine hydrochloride (or C-390) and an agent capable of inhibiting Pseudomonas aeruginosa bacteria. The invention also relates to a culture medium suitable for implementing this process for the enrichment and selective culture of mycobacteria.

First claim

The invention claimed is:

1. A process for the enrichment and selective culture of mycobacteria contained in a biological sample, the process comprising:
inoculating all or part of the biological sample in/on a culture medium comprising:
a nutritive composition reproducing that of a Middlebrook medium supplemented with at least:

glycerol,
a supplement based on oleic acid, albumin, dextrose, and catalase (an OADC supplement), and
a yeast extract;
9-chloro-9-(4′-diethylamino)phenyl-9,10-dihydro-10-phenylacridine hydrochloride (C-390); and
an agent capable of inhibiting Pseudomonas aeruginosa bacteria.

2. The process as claimed in claim 1 , wherein the C-390 concentration of the culture medium is between 2 mg/l and 1000 mg/l.

3. The process as claimed in claim 1 , wherein the agent capable of inhibiting Pseudomonas aeruginosa bacteria is chosen from: colistin methanesulfonate, fosfomycin, nalidixic acid, aztreonam, cefsulodine and mangrolide A.

4. The process as claimed in claim 1 , wherein the culture medium also comprises an agent capable of inhibiting Burkholderia cepacia bacteria.

5. The process as claimed in claim 1 , wherein the culture medium also comprises at least one selective agent chosen from: amphotericin B, nalidixic acid, vancomycin, colistin methanesulfonate, fosfomycin and malachite green.

6. The process as claimed in claim 5 , wherein the culture medium comprises, at least one selective agent chosen from:
amphotericin B, at a concentration of between 2 mg/l and 50 mg/1;
colistin methanesulfonate, at a concentration of between 15 mg/l and 150 mg/1; and
fosfomycin, at a concentration of between 200 mg/l and 2000 mg/l.

7. The process as claimed in claim 6 , wherein the culture medium also comprises, in addition to the fosfomycin, glucose-6-phosphate, at a concentration of between 10 mg/l and 50 mg/l.

8. A process for the enrichment and selective culture of mycobacteria contained in a biological sample, the process comprising:
inoculating all or part of the biological sample in/on a culture medium comprising:
a nutritive composition reproducing that of a Middlebrook medium, supplemented with:

glycerol, at a concentration of between 2 ml/l and 20 ml/l,
an OADC supplement, at a concentration of between 50 ml/l and 200 ml/l, and
a yeast extract, at a concentration of between 0.1 g/l and 20 g/l;
9-chloro-9-(4′-diethylamino)phenyl-9,10-dihydro-10-phenylacridine hydrochloride (C-390); and
an agent capable of inhibiting Pseudomonas aeruginosa bacteria.

9. A process for the enrichment and selective culture of mycobacteria contained in a biological sample, the process comprising:
inoculating all or part of the biological sample in/on a culture medium comprising:
a nutritive composition reproducing that of a Löwenstein-Jensen medium;
C-390; and
an agent capable of inhibiting Pseudomonas aeruginosa bacteria.

10. The process as claimed in claim 9 , wherein the C-390 concentration of the culture medium is between 2 mg/l and 1000 mg/l.

11. The process as claimed in claim 9 , wherein the agent capable of inhibiting Pseudomonas aeruginosa bacteria is chosen from: colistin methanesulfonate, fosfomycin, nalidixic acid, aztreonam, cefsulodine and mangrolide A.

12. The process as claimed in claim 9 , wherein the culture medium also comprises an agent capable of inhibiting Burkholderia cepacia bacteria.

13. The process as claimed in claim 9 , wherein the culture medium also comprises at least one selective agent chosen from: amphotericin B, nalidixic acid, vancomycin, colistin methanesulfonate, fosfomycin and malachite green.

Assignees

• Biomerieux Sa

Inventors

• Orenga Sylvain
• Perry Audrey
• Perry John
• Preece Clair

Classifications

• C12Q1/045 Primary

  Culture media therefor · CPC title

• C12N1/20 Primary

  Bacteria; Culture media therefor · CPC title

• G01N2333/35

  from Mycobacteriaceae (F) · 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

• C12N1/02

  Separating microorganisms from their culture media · CPC title