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Growth medium for the detection of microorganisms by fluorescence allying a fluorogenic substrate and a pH-sensitive fluorophore

A technology of growth medium and fluorescent substrate, which is used in the field of growth medium for fluorescence detection of microorganisms by combining fluorescent substrates and pH-sensitive fluorophores, and can solve problems such as inability to distinguish microorganisms and inability to determine relative proportions.

Active Publication Date: 2012-10-10
MILLIPORE CORP
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  • Abstract
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  • Application Information

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Problems solved by technology

As such, they cannot distinguish between microorganisms present in the same growth medium, nor can they determine their relative proportions

Method used

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  • Growth medium for the detection of microorganisms by fluorescence allying a fluorogenic substrate and a pH-sensitive fluorophore
  • Growth medium for the detection of microorganisms by fluorescence allying a fluorogenic substrate and a pH-sensitive fluorophore
  • Growth medium for the detection of microorganisms by fluorescence allying a fluorogenic substrate and a pH-sensitive fluorophore

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Embodiment Construction

[0122] 1 Preparation of growth medium

[0123] 100 mL of liquid TSB (Biomerieux) medium was prepared and sterilized according to the manufacturer's recommendations. Then, the following solutions of pH-sensitive fluorophores and fluorogenic substrates were added under aseptic conditions to the TSB medium according to each combination described below.

[0124] pH-sensitive fluorophores:

[0125] HTPS:

[0126] - 30 μL of 10 mM stock solution of 8-hydroxypyrene 1,3,6-trisulfonic acid, or

[0127] CF:

[0128] - 20 μL of a sterile stock solution of 100 μM 5,6-carboxyfluorescein;

[0129] Fluorescent substrate:

[0130] Aldols:

[0131] - 200 μL of a sterile mixture of Aldol derivatives (Aldol 470 phosphate, Aldol 470 ester, aldol 455β-D-glucopyranoside, aldol 455β-D-galactopyranoside), where the stock concentration of each derivative 25mg / mL; or

[0132] 4-MU:

[0133] - 200 μL of 4-methylumbelliferyl derivatives (4-MU phosphate, 4-MUβ-D-glucopyranoside, 4-MUα-D-glucopy...

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Abstract

The invention relates to a growth medium allying a fluorogenic substrate and a pH-sensitive fluorophore, in particular the combination of 4- methylumbelliferone (4-MU) and derivatives of fluorescein. This growth medium is used for the detection by fluorescence of microorganisms by coupling the fluorescence measurements relating to the pH-sensitive fluorophor and the fluorescence measurements relating to the activation of the fluorogenic substrate(s) by the microorganisms.

Description

technical field [0001] The present invention relates to methods and devices for testing the sterility of aqueous or gaseous fluids used in particular in the manufacture of pharmaceutical products and devices. [0002] In particular, the present invention relates to a growth medium comprising a fluorogenic substrate and a pH-sensitive fluorophore, in particular a combination of 4-methylumbelliferone (4-MU) and a fluorescein derivative, for use by fluorescent , in particular for the detection of microorganisms by combining fluorescence detection of pH-sensitive fluorophores with fluorescence detection of fluorescent substrates activated by the microorganisms. Background technique [0003] The pharmaceutical industry is one of those areas where sterility restrictions are very high, forcing continuous monitoring of product quality and all activities related to these products. [0004] For microbial contamination, tests are performed using microbiological methods in accordance w...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/04C12N1/20
CPCC12Q1/045
Inventor F・马克
Owner MILLIPORE CORP
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