Compositions and methods for disease diagnosis using single cell analysis
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Publication Number
US-11061018-B2
Publication Date
2021-07-13
Expiration Date
Abstract
Certain embodiments of the invention are directed to evaluating and identifying cells by recording and interpreting a time-dependent signal produced by unique cell respiration and permeability attributes of isolated viable cells.
Core Innovation
The invention relates to detecting and characterizing microbes in a sample by isolating microbe cells and separating non-microbe cells, viruses, proteins, and nucleic acids to form an isolated sample. The isolated sample is divided into at least two sample portions and placed in different growth media, and each sample portion includes a reporter to form a first and second sample mixture.
The first and second sample mixtures are partitioned so that some partitions have at most one microbial cell or microbial cell aggregate per partition. The partitions are incubated over time at a specific temperature or temperatures while the reporter interacts with the microbial cell or microbial cell aggregate in the partition.
During incubation, optical characteristics are monitored to measure the amount of optical signal produced by interaction of the reporter with the microbial cell or microbial cell aggregate. For each partition, an optical signal waveform is constructed, resulting in a partition waveform.
Microbes are detected and characterized by comparing the first and second partition waveforms between the different growth-media portions and assessing microbes based on the partition waveform comparison.
Claims Coverage
Independent claim clm-00001 defines a complete workflow with six inventive features: microbe isolation, dual growth-medium portions with a reporter, partitioning with at most one microbial cell or aggregate in some partitions, time incubation, reporter-driven optical monitoring to form waveforms, and waveform comparison between portions for detection and characterization.
Microbe isolation into an isolated sample
The method isolates microbes from a sample by separating non-microbe cells, viruses, proteins, and nucleic acids from microbe cells to form an isolated sample.
Dual growth-medium sample portions with a reporter
The method divides the sample into at least two sample portions and suspends a first portion in a first growth medium and a second portion in a second growth medium, with each portion including a reporter, forming a first and a second sample mixture.
Partitioning with at most one microbial cell or aggregate
The method partitions the first and second sample mixtures to form first and second sample partitions, where some partitions have at most one microbial cell or microbial cell aggregate per partition.
Time incubation of partitions at specific temperature
The method incubates the partitions over time at a specific temperature or temperatures.
Reporter-driven optical signal waveform construction
The method monitors optical characteristics of the partitions during incubation, including the amount of optical signal produced by interaction of the reporter with the microbial cell or microbial cell aggregate, and constructs an optical signal waveform for each partition to obtain a partition waveform.
Waveform comparison between growth media for detection and characterization
The method compares partition waveforms between the first and second sample partition waveforms and assesses microbes based on the partition waveform comparison, detecting and characterizing microbes based on the comparison of the first and second waveforms from each sample portion.
Overall claim coverage centers on detecting and characterizing microbes by combining isolation, reporter-containing sample portions in different growth media, partitioning into partitions with at most one microbial cell or aggregate in some partitions, time incubation, construction of reporter-driven optical signal waveforms for each partition, and comparison of waveforms between the different sample portions to assess microbes.
Stated Advantages
Not explicitly described in patent.
Documented Applications
Not explicitly described in patent.
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