Enzymatic substrates for detecting Pseudomonas aeruginas
AssigneeBiomerieux
InventorsMireille Desmonceaux, Daniel Monget
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Abstract
An enzyme substrate which includes a target portion and a marker portion, hydrolysis of the substrate leading to separation of the target portion from the marker portion, and the target portion being specific for the enzyme activity being assayed. A formulation containing at least one such substrate, and a method for detecting the bacterial species Pseudomonas aeruginosa is also disclosed. The target portion of the substrate is specific for β-alanine aminopeptidase activity and the marker portion is a compound which reveals whether a hydrolysis reaction has taken place or not. The invention is particularly applicable in the field of diagnosis.
Claims15
An enzyme substrate comprising a target portion and a marker portion, said target portion and marker portion being separable by hydrolysis, said target portion being specific for the enzyme activity being assayed, such that said target portion is acted upon by the enzyme β-alanine aminopeptidase derived from at least one Pseudomonas aeruginosa microorganism, and the marker portion is a molecule which reveals whether a hydrolysis reaction has taken place or not, wherein said substrate has the formula:
- H2N—CH2—CH2—CO—NH—R,
- in which H2N—CH2—CH2— CO is the target portion and —NH—R is the marker portion of the substrate.
A formulation for detecting at least one strain and/or species of microorganism comprising
- at least one enzyme substrate comprising a target portion and a marker portion, said target portion and marker portion being separable by hydrolysis, said target portion being specific for the enzyme activity being assayed, such that said target portion is acted upon by the enzyme β-alanine aminopeptidase derived from at least one Pseudomonas aeruginosa microorganism, and the marker portion is a molecule which reveals whether a hydrolysis reaction has taken place or not, wherein said substrate has the formula:
- H2N—CH2—CH2—CO—NH—R,
- in which H2N—CH2—CH2—CO— is the target portion and —NH—R is the marker portion of the substrate, and
- a culture medium.
A formulation for detecting at least one strain and one species of microorganism, or at least two strains or two species of microorganism, comprising
- at least one enzyme substrate comprising a target portion and a marker portion, said target portion and marker portion being separable by hydrolysis, said target portion being specific for the enzyme activity being assayed, such that said target portion is acted upon by the enzyme β-alanine aminopeptidase derived from at least one Pseudomonas aeruginosa microorganism, and the marker portion is a molecule which reveals whether a hydrolysis reaction has taken place or not, wherein said substrate has the formula:
- H2N—CH2—CH2—CO—NH—R,
- in which H2N—CH2—CH2—CO— is the target portion and —NH—R is the marker portion of the substrate,
- a culture medium, and
- at least one other substrate.
A method for detection of Pseudomonas aeruginosa, comprising:
- contacting at least one substrate capable of detecting β-alanine aminopeptidase enzyme activity with a sample suspected of containing at least one Pseudomonas aeruginosa microorganism, and
- monitoring for the appearance of a colored and/or fluorescent signal from said sample and which correlates to said β-alanine aminopeptidase enzyme activity.
- wherein said substrate has the formula:
- H2N—CH2—CH2—CO—NH—R,
- in which H2N—CH2—CH2—CO— is the target portion and —NH—R is the marker portion of the substrate.
A method for detection of Pseudomonas aeruginosa, comprising:
- contacting at least one substrate capable of detecting β-alanine aminopeptidase enzyme activity with a sample suspected of containing at least one Pseudomonas aeruginosa microorganism,
- contacting at least one substrate capable of detecting the activity of an enzyme other than β-alanine aminopeptidase with a sample suspected of containing at least one microorganism other than Pseudomonas aeruginosa, this other enzyme activity making it possible to distinguish Pseudomonas aeruginosa from said other microorganism, and
- monitoring for the appearance of a colored and/or fluorescent signal from said sample and which correlates to said β-alanine peptidase enzyme activity.
- wherein said substrate has the formula:
- H2N—CH2—CH2—CO—NH—R,
- in which H2N—CH2—CH2—CO— is the target Portion and —NH—R is the marker portion of the substrate.
Dependencies are inferred from the claim text.