Electrochemiluminescence assay
OwnerMark T. Martininventor
Life of this patent
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Abstract
A rapid single step assay suitable for the detection or quantification of .beta.-lactam antibiotics and .beta.-lactamases. The assay can be performed directly on samples of food, such as milk and meat, blood or serum and is useful in determining the suitability of a particular antibiotic in treating a particular bacterial infection and in diagnosis of a bacterial infection. The assay is also useful in determining and quantifying .beta.-lactam antibiotic resistance. The assay can be performed on an IGEN Origen.sup.R Analyzer.
Claims31
A kit for measuring .beta.-lactam antibiotics or .beta.-lactamases comprising premeasured amounts of a bidentate aromatic heterocyclic nitrogen-containing ligand of ruthenium or osmium reagent and a .beta.-lactam or lactamase standard wherein the premeasured amounts are sufficient to perform a single sample measurement.
A method for measuring the presence of .beta.-lactamase comprising,
- (a) contacting a sample suspected of containing .beta.-lactamase with a .beta.-lactam and a bidentate aromatic heterocyclic nitrogen-containing ligand of ruthenium or osmium reagent;
- (b) subjecting the contacted sample of step (a) to conditions which result in electrochemiluminesence;
- (c) measuring the resulting electrochemiluminescence to obtain a reading;
- (d) incubating the mixture;
- (e) subjecting the incubated mixture of step (d) to conditions which result in electrochemiluminesence;
- and
- (f) measuring the resulting chemiluminescence and comparing it to the previously measured value to determine the difference, if any, in the measured values thereby indicating the presence of .beta.-lactamase.
A method for measuring the presence of .beta.-lactam comprising,
- (a) contacting a sample containing .beta.-lactam with a .beta.-lactamase and a bidentate aromatic heterocyclic nitrogen-containing ligand of ruthenium or osmium reagent;
- (b) subjecting the contacted sample of step (a) to conditions which result in electrochemiluminesence;
- (c) measuring the resulting electrochemiluminescence to obtain a reading;
- (d) incubating the mixture;
- (e) subjecting the incubated mixture of step (d) to conditions which result in electrochemiluminesence;
- (f) measuring the resulting chemiluminescence and comparing it to the previously measured value to determine the presence of a difference in measured values;
- and
- (g) comparing the difference with a standard value to determine the quantitative amount of .beta.-lactam present in the sample.
A method for identifying .beta.-lactams comprising,
- (a) preparing a series of samples containing a .beta.-lactam of interest from a sample source;
- (b) contacting the series of samples containing the .beta.-lactam of interest with a bidentate aromatic heterocyclic nitrogen-containing ligand of ruthenium or osmium reagent and a series of different .beta.-lactamases;
- (c) subjecting each of contacted samples of step (b) to conditions which result in electrochemiluminesence;
- (d) separately measuring the resulting electrochemiluminescence for each sample of the series to obtain a series of base-line readings;
- (e) incubating the series of samples of step (d);
- (f) subjecting the incubated samples of step (e) to conditions which result in electrochemiluminesence;
- (g) separately measuring the resulting chemiluminescence for each incubated sample and comparing it to the base-line reading for the corresponding sample in the series to determine the presence of a difference in the measured values;
- (h) generating a spectrum of differences in measured values for the series;
- and
- (i) comparing the generated spectrum with a standard spectrum to identify the .beta.-lactam present in the source material.
A method for identifying bacteria comprising,
- (a) preparing a series of samples containing the bacteria of interest from a sample material;
- (b) contacting a series of samples suspected of containing the same bacteria with a bidentate aromatic heterocyclic nitrogen-containing ligand of ruthenium or osmium reagent and a series of different .beta.-lactams;
- (c) subjecting each of the contacted sample of step (b) to conditions which result in electrochemiluminesence;
- (d) measuring the resulting electrochemiluminescence to obtain a base-line reading for each of the samples in the series;
- (e) incubating the series of samples of step (d);
- (f) subjecting the incubated samples of step (e) to conditions which result in electrochemiluminesence;
- (g) measuring the resulting chemiluminescence for each sample in the series and comparing it to the corresponding base-line value for the same sample in the series to determine the presence of a difference in measured values;
- (h) generating a spectrum of differences in measured values for the series;
- and
- (i) comparing the generated spectrum with a spectrum standard to identify the bacteria in the source material.
Dependencies are inferred from the claim text.