Detection of bacteria using bacteriophage
Inventors
Kellum, John Alston • HEMPEL, JOHN D. • EDGAR, ROBERT HUGH • VIATOR, JOHN ANDREW
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Assignees
University of Pittsburgh • Duquesne University of the Holy Spirit
Member
Nanomedicine Manufacturing Lab, Duquesne UniversityNANOMEDICINE MANUFACTURING LABORATORY
Nanomedicines produced at NML include colloidal nanosystems for molecular imaging (magnetic resonance imaging (MRI) and near-infrared fluorescence (NIRF) imaging), targeted and local drug delivery, and imaging-supported drug delivery and theranostic nanomedicines and biomaterials. These products can be delivered locally, parenterally, or implanted into body cavities or wounds. Nanotechnology-based therapeutics are typically presented with high costs and challenging quality control, representing critical barriers to future clinical translation. In contrast, the offeror NML efforts over the past decade produced cost-effective, robust, and scalable manufacturing methods for nanomedicines with a high level of quality control by utilizing Quality-by-Design (QbD) approaches. Specifically, the application of QbD to nanomedicine manufacturing and quality control led to several firsts: 1) the first imaging-supported pain nanomedicine for trauma and surgical pain; 2) the first oxygen carrier with embedded imaging agents for real-time in line tracking during organ/limb preservation; 3) the first successful longitudinal immunomonitoring in non-human primates and porcine models using clinical grade imagers; 4) demonstrated nerve injury recovery following trauma by local nanomedicine implantation in rodents. NML also designs and produces biocompatible and multi-drug delivery hydrogels and biomaterials for multitude of applications, from supporting neuroregeneration to local immunosuppression and wound healing. Furthermore, NML successfully scaled up their laboratory protocols to produce >2L of nanoparticles/batch and evaluate them in human limb trials for oxygen delivery. The work in these areas has been supported by USAF and CDMRP contracts, which are highly collaborative and involve partners across academia, industry and Government. NML is currently funded by CDMRP and ARPA H.
Founded in 1878, Duquesne University is consistently ranked among the nation's top Catholic universities for its award-winning faculty and horizon-expanding education. Research happens in all fields across the University, from the humanities and sciences to health-related fields and business. This research is supported by the federal and state governments, foundations, and corporate partners. Duquesne's Pittsburgh location connects researchers of all kinds to a knowledge economy powered by large tech, medical, energy, and industrial sectors.
NANOMEDICINE MANUFACTURING LABORATORY Nanomedicines produced at NML include colloidal nanosystems for molecular imaging (magnetic resonance imaging (MRI) and near-infrared fluorescence (NIRF) imaging), targeted and local drug delivery, and imaging-supported drug delivery and theranostic nanomedicines and biomaterials. These products can be delivered locally, parenterally, or implanted into body cavities or wounds. Nanotechnology-based therapeutics are typically presented with high costs and challenging quality control, representing critical barriers to future clinical translation. In contrast, the offeror NML efforts over the past decade produced cost-effective, robust, and scalable manufacturing methods for nanomedicines with a high level of quality control by utilizing Quality-by-Design (QbD) approaches. Specifically, the application of QbD to nanomedicine manufacturing and quality control led to several firsts: 1) the first imaging-supported pain nanomedicine for trauma and surgical pain; 2) the first oxygen carrier with embedded imaging agents for real-time in line tracking during organ/limb preservation; 3) the first successful longitudinal immunomonitoring in non-human primates and porcine models using clinical grade imagers; 4) demonstrated nerve injury recovery following trauma by local nanomedicine implantation in rodents. NML also designs and produces biocompatible and multi-drug delivery hydrogels and biomaterials for multitude of applications, from supporting neuroregeneration to local immunosuppression and wound healing. Furthermore, NML successfully scaled up their laboratory protocols to produce >2L of nanoparticles/batch and evaluate them in human limb trials for oxygen delivery. The work in these areas has been supported by USAF and CDMRP contracts, which are highly collaborative and involve partners across academia, industry and Government. NML is currently funded by CDMRP and ARPA H. Founded in 1878, Duquesne University is consistently ranked among the nation's top Catholic universities for its award-winning faculty and horizon-expanding education. Research happens in all fields across the University, from the humanities and sciences to health-related fields and business. This research is supported by the federal and state governments, foundations, and corporate partners. Duquesne's Pittsburgh location connects researchers of all kinds to a knowledge economy powered by large tech, medical, energy, and industrial sectors.
Publication Number
US-10544443-B2
Publication Date
2020-01-28
Expiration Date
2036-07-29
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 and system for detecting a species, strain, or type of bacteria by mixing a labeled bacteriophage—with a detectable label—into a bacterial culture. The labeled bacteriophage selectively binds to its target bacteria, and a detection system, which can include a photoacoustic cell, is used to detect the presence of labeled bacteriophage bound to bacteria. The method may involve removing unbound labeled bacteriophage prior to detection, and allows for identification and quantification of the targeted bacteria.
The method can be extended by using a plurality of different labeled bacteriophages, each selective for a different bacterial species, strain, or type. Each bacteriophage is labeled with a detectably distinct tag, for example, one identifiable at a unique wavelength. Thus, the detection system can indicate the presence of multiple specific bacterial types within a single sample. The method includes the use of various photoacoustic-compatible labels and is adaptable to other detection technologies, such as flow cytometry or electron microscopy.
The problem addressed by the invention is the slow, inaccurate, and limited nature of current bacterial detection methods, which require 48 hours or longer and are unable to accurately quantify detected bacteria or detect bacteria that do not grow readily on agar plates. The disclosed method enables more rapid, quantitative, and specific bacterial detection.
Claims Coverage
There is one independent claim, focusing on the method of detecting bacteria using labeled bacteriophage and a photoacoustic detection system.
Method of detecting bacteria using labeled bacteriophage and photoacoustic cell
A method comprises: - Mixing a labeled bacteriophage, which includes a label detectable via a detection system comprising a photoacoustic cell, with a biological sample. - The labeled bacteriophage must be active to selectively bind with the species, strain, or type of bacteria intended for detection. - Using the detection system (with a photoacoustic cell) to determine the presence of the labeled bacteriophage bound to the targeted species, strain, or type of bacteria in the sample.
The coverage centers on the use of labeled bacteriophage, photoacoustic detection, and selective binding for determining if a sample contains a specific species, strain, or type of bacteria.
Stated Advantages
The method provides a quicker and quantitative assay for bacterial detection, delivering results in hours instead of days.
It allows identification and quantification of specific bacterial species, strains, or types in a sample.
The technique is applicable to bacteria that are hard or impossible to grow in culture.
Photoacoustic detection offers increased detection rates and specificity at a reduced cost compared to other detection systems.
Earlier detection using this method enables earlier treatment with targeted antibiotics.
The assay supports single-step estimation of bacterial concentration, distinguishing pathogen from flora.
Documented Applications
Rapid detection and quantification of bacterial contamination in clinical samples, such as blood or sputum, in hospital and medical laboratory settings.
Guiding antibiotic selection for infections based on identification of causative bacterial species, including detection of bacteria not culturable by standard methods.
Facilitating early diagnosis and treatment of pneumonia and differentiating bacterial from viral infections in clinical scenarios.
Supporting detection of multi-drug resistant pathogens and minimizing unnecessary use of broad-spectrum antibiotics.
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