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US 9,175,329 B2Granted
Methods for antimicrobial resistance determination
AssigneeBio Merieux
InventorsJohn Walsh, Jones Hyman
Life of this patent
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Abstract
The present invention relates to methods and systems for determining the antibiotic-resistance status of microorganisms. The invention further provides methods for determining the antibiotic-resistance status of microorganisms in situ within a single system.
Claims20
A method for determining the antibiotic-resistance status of a microorganism, comprising:
- (a) contacting the microorganism with a resistance-determining affinity ligand under conditions whereby a microorganism/resistance-determining affinity ligand complex can be formed;
- (b) separating microorganism/resistance-determining affinity ligand complex formed in (a) from unbound resistance-determining affinity ligand;
- (c) interrogating and determining the amount of resistance-determining affinity ligand bound to the microorganism in the microorganism/resistance-determining affinity ligand complex;
- and
- (d) comparing the amount of resistance-determining affinity ligand bound to the microorganism in the microorganism/resistance-determining affinity ligand complex to the amount of resistance-determining affinity ligand bound by a known antibiotic-sensitive or antibiotic-resistant strain of the same microorganism or a population of known antibiotic-sensitive or antibiotic-resistant strains;
- wherein one or more of steps (a), (b), (c), and/or (d) are carried out in a hermetically sealed container;
- and wherein if the microorganism in the microorganism/resistance-determining affinity ligand complex binds a different amount of resistance-determining affinity ligand than is bound by the antibiotic-sensitive microorganism or the microorganism in the microorganism/resistance-determining affinity ligand complex binds the same amount of resistance-determining affinity ligand as is bound by the antibiotic-resistant microorganism, the microorganism is identified as antibiotic-resistant.
4 more elements
A method for determining the antibiotic-resistance status of a microorganism, comprising:
- (a) obtaining a test sample known to contain or that may contain microorganisms;
- (b) contacting the microorganism in the test sample with a resistance-determining affinity ligand under conditions whereby a microorganism/resistance-determining affinity ligand complex can be formed;
- (c) optionally adding one or more fluorescent dyes that measure cell metabolism or membrane integrity;
- (d) optionally selectively lysing any non-microbial cells that may be present in the sample to produce a lysed sample;
- (e) separating the microorganisms from other components of said test sample, or said lysed sample, to form a pellet of microorganisms;
- (f) spectroscopically interrogating the pellet to produce measurements of the microorganism;
- (g) determining the antibiotic-resistance status of the microorganisms in the test sample by comparison of the measurements with measurements taken, or predicted, for antibiotic-resistant and/or antibiotic-susceptible microorganisms of the same species;
- and
- wherein one or more of steps (b), (c), (d), (e) and/or (f) are carried out in a hermetically sealed container.
6 more elements
Dependencies are inferred from the claim text.