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US 11,634,782 B2Granted
Quantification of microorganisms in samples and methods of determining quantification conditions thereof
AssigneeHygiena, LLC
InventorTyler Stephens
Life of this patent
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Abstract
This disclosure relates to methods of determining quantification conditions for a microorganism and methods of quantifying microorganism concentration in a sample.
Claims27
A method of determining quantification conditions for a microorganism in a sample comprising:
- (a) inoculating a plurality of samples from a sample lot with logarithmic serial dilutions of a microorganism in a quantification medium;
- (b) incubating the plurality of samples at one or more temperatures for a time period sufficient to detect the microorganism in the plurality of samples by a detection assay;
- (c) removing aliquots of the plurality of samples at a plurality of time points and at one or more temperatures of the incubation;
- (d) detecting a target nucleic acid sequence of the microorganism in each aliquot by the detection assay;
- and
- (e) determining a detection value at each time point and inoculation level to produce a linear best fit curve for the microorganism in the sample lot.
3 more elements
A method of quantifying microorganism concentration in a sample comprising:
- (a) collecting a test sample from a test sample lot;
- (b) incubating the test sample (i) in a quantification medium (ii) for a pre-determined quantification time, (iii) at a pre-determined dilution ratio, and (iv) at a pre-determined temperature, wherein (i)-(iv) correlate to a pre-determined linear best fit curve for the microorganism from the test sample lot;
- (c) detecting presence or absence of a target nucleic acid sequence of the microorganism in the test sample by a detection assay;
- and
- (d) comparing the results of the detection assay with the pre-determined linear best fit curve to quantify an amount of microorganism in the test sample.
2 more elements
A method of quantifying microorganism concentration in a sample comprising:
- (a) inoculating a plurality of samples from a sample lot with logarithmic serial dilutions of a microorganism in a quantification medium;
- (b) incubating the plurality of samples in the quantification medium at one or more temperatures for a time period sufficient to detect the microorganism in the plurality of samples by a detection assay;
- (c) removing aliquots of the plurality of samples at a plurality of time points of the incubation;
- (d) detecting a target nucleic acid sequence of the microorganism in each aliquots by the detection assay;
- (e) determining a detection value at each time point and inoculation level to produce a linear best fit curve for the microorganism in the sample lot;
- (f) collecting a test sample from a test sample lot, wherein the test sample is of the same type as the sample lot;
- (g) incubating the test sample in the quantification medium (i) for a pre-determined quantification time, (ii) at a pre-determined dilution ratio, and (iii) at a pre-determined temperature, wherein (i)-(iii) correlate to the linear best fit curve for the microorganism in the sample lot;
- (h) detecting presence or absence of a test target nucleic acid sequence of the microorganism in the test sample by the detection assay;
- and
- (i) comparing the results of the detection assay of step (h) with the linear best fit curve to quantify an amount of microorganism in the test sample.
7 more elements
Dependencies are inferred from the claim text.