Detection of bacteria using bacteriophage
Inventors
Kellum, John Alston • HEMPEL, JOHN D. • EDGAR, ROBERT HUGH • VIATOR, JOHN ANDREW
Assignees
University of Pittsburgh • Duquesne University of the Holy Spirit
Publication Number
US-11629370-B2
Publication Date
2023-04-18
Expiration Date
2036-07-29
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Abstract
A method of detecting a species, strain or type of bacteria includes mixing a labeled bacteriophage including a label that is detectible via a detection system with a bacterial culture including the species, strain or type of bacteria to which the labeled bacteriophage selectively binds and using the detection system to detect the labeled bacteriophage bound to the species, strain or type of bacteria.
Core Innovation
The invention provides a method for detecting or determining a species, strain, or type of bacteria by mixing a labeled bacteriophage, which includes a label detectable through a detection system, with a bacterial culture containing the target bacteria. The labeled bacteriophage selectively binds to the target bacteria, and the detection system is used to identify the labeled bacteriophage bound to the bacteria. The system may incorporate a step to remove unbound labeled bacteriophage, and the detection system can include a photoacoustic cell or photoacoustic flowmetry system for detection and quantification.
The background highlights that current bacterial detection methods in samples such as body fluids are slow, requiring 48 hours or more, lack accurate quantification, and are limited to bacteria that can grow readily on agar plates. The present invention addresses these limitations by enabling a faster, quantitative assay. By labeling bacteriophages with detectable tags and using detection systems such as photoacoustic cells, the presence and type of bacteria can be rapidly and accurately determined, allowing timely and specific clinical interventions.
In specific embodiments, a plurality of labeled bacteriophages, each selectively binding to different species, strains, or types of bacteria and each with a distinct detectable label, can be mixed into the sample. The detection system discerns which labeled bacteriophage has bound, thus enabling simultaneous identification of multiple bacterial types. The method can identify and quantify bacteria, with detection enhanced by the spatial sequestration of the labeled phage when bound, and signal clarity improved by using detection system energy levels low enough to exclude unbound labeled bacteriophages.
Claims Coverage
The patent contains one independent claim covering a method of determining if a sample contains a species, strain, or type of bacteria, with several main inventive features.
Bacteriophage-based detection using photoacoustic signals
The method involves mixing a labeled bacteriophage, which includes a label detectable via a detection system, with a sample. The labeled bacteriophage selectively binds with the species, strain, or type of bacteria. Light energy is applied to the sample to generate photoacoustic waves, and these generated photoacoustic waves are measured to determine the presence of the labeled bacteriophage bound to the target bacteria.
The inventive coverage focuses on using labeled bacteriophages and photoacoustic detection to rapidly and specifically identify bacteria in a sample.
Stated Advantages
The method provides a much faster assay for bacterial detection, reducing the time to obtain results from days to hours.
It allows for accurate quantification and identification of bacterial species, strains, or types, including pathogens that are difficult or impossible to grow in culture.
The assay can be used in clinical settings to support earlier and targeted decisions for antibiotic treatment, improving patient management.
Photoacoustic detection offers increased sensitivity and specificity over other systems such as flow cytometry or electron microscopy, often at reduced cost.
The approach can estimate bacterial counts rapidly in a single step, avoiding slow colony counting methods.
Documented Applications
Clinical detection of bacterial pathogens in patient samples (e.g., blood, sputum) to facilitate targeted antibiotic therapy.
Rapid bacterial typing in hospital and medical laboratory settings to detect and quantify bacterial contamination.
Simultaneous detection of multiple bacteria by using different labeled bacteriophages in a sample.
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