Microbial susceptibility assay using 7-N-(aminoacyl)-7-amido-4-methylcoumarins
OwnerAnthony H. Carr, Robert A. Badley +2inventors
InventorsAnthony H. Carr, Robert A. Badley, Ian Jobling, Thomas J. Sands
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Abstract
A method of determining the susceptibility of microorganisms to an antimicrobial substance wherein the microorganisms are cultured in the presence of the antimicrobial substance, and growth or lack of growth is assessed by detecting a fluorescent product resulting from the enzymatic hydrolysis of a fluorogenic substrate. The fluorogenic substrates include 7-N-(aminoacyl)-7-amido-4-methylcoumarin, 7-N-(alanyl)-7-amido-4-methylcoumarin, or a combination of 7-N-(alanyl)-7-amido-4-methylcoumarin and 4-methyl umbelliferyl phosphate.
Claims10
In a method of determining the susceptibility of microorganisms to an antimicrobial substance, comprising culturing the microorganisms in the presence of an antimicrobial substance, contacting the microorganisms with a fluorogenic substrate which is enzymatically hydrolyzable to yield a fluorescent product, and determining growth by measuring the fluorescent product yielded by the hydrolysis of the fluorescent substrate, wherein the improvement comprises using as said fluorogenic substrate a 7-N-(aminoacyl)-7-amido-4-methylcoumarin and exposing the microorganisms to the fluorescent substrate throughout culturing.
In a method of determining the susceptibility of microorganisms to an antimicrobial substance, comprising culturing the microorganisms in the presence of an antimicrobial substance, contacting the microorganisms with a fluorogenic substrate which is enzymatically hydrolyzable to yield a fluorescent product, and determining growth by measuring the fluorescent product yielded by the hydrolysis of the fluorescent substrate, wherein the improvement comprises using as said fluorogenic substrate a 7-N-(alanyl)-7-amido-4-methylcoumarin.
In a method of determining the susceptibility of microorganisms to an antimicrobial substance, comprising culturing the microorganisms in the presence of an antimicrobial substance, contacting the microorganisms with a fluorogenic substrate which is enzymatically hydrolyzable to yield a fluorescent product, and determining growth by measuring the fluorescent product yielded by the hydrolysis of the fluorescent substrate, wherein the improvement comprises using as said fluorogenic substrate a combination of 7-N-(aminoacyl)-7-amido-4-methylcoumarin and 4-methyl umbelliferyl phosphate.
Dependencies are inferred from the claim text.