Apparatus and method for detecting microscopic organisms using bacteriophage
Inventors
Assignees
Interested in licensing this patent?
MTEC can help explore whether this patent might be available for licensing for your application.
Publication Number
US-8092990-B2
Publication Date
2012-01-10
Expiration Date
Abstract
Methods and apparatus are provided for detection of microorganisms in a sample. Methods and apparatus of the invention are based on the specificity that phage, for example bacteriophage, have for target microorganisms, for example bacterium. Typically, phage adsorption to target microorganisms act as signal, or a signal target, for the presence of the target microorganism. Typically, the phage are labeled with a detectable signal. Apparatus of the invention are directed toward concentrating the phage adsorbed microorganisms at a predetermined site for flag dependent observation.
Core Innovation
The invention provides a method of determining the presence of a target microorganism in a sample by combining the sample with an amount of flagged phage capable of binding to an outside surface of the target microorganism. The flagged phage binds to the outside surface to create a flag labeled target microorganism, and conditions and an amount of time are provided sufficient for the flagged phage to bind to the outside surface.
The method then assays the phage exposed sample to detect the presence of the flag labeled microorganism without detecting phage amplified within the target microorganism. Presence of the flag bound to the outside surface indicates presence of the microorganism, and detection is supported by concentrating the flag labeled microorganism to enhance capacity to detect the flag labeled microorganisms.
Concentrating is carried out on a flow strip device having one or more immobilization zones, where the flag labeled microorganisms are captured and concentrated within the one or more immobilization zones. In preferred embodiments, the flagged phage is conjugated to visible or colorimetric flags such as colloidal gold, or alternatively fluorescent tags, enabling visual readout at capture locations on the detection devices.
Claims Coverage
The independent claim provides an integrated detection approach with two principal inventive features: using flagged phage binding to an outside surface of the target microorganism with detection that does not rely on phage amplified within the microorganism, and enhancing detection by concentrating the flag labeled microorganisms on a flow strip device with immobilization zones.
Outside surface binding flagged phage for detecting target microorganism
Combining the sample with an amount of flagged phage capable of binding to an outside surface of the target microorganism to create a flag labeled target microorganism, providing conditions and an amount of time sufficient for the flagged phage to bind to the outside surface, and assaying the phage exposed sample to detect the presence of the flag labeled microorganism without detecting phage amplified within the target microorganism, wherein presence of the flag bound to the outside surface indicates presence of the microorganism.
Concentrating flag labeled microorganisms on immobilization zones within a flow strip device
Detecting the presence of the flag labeled microorganism by concentrating the flag labeled microorganisms to enhance the capacity to detect the flag labeled microorganisms, wherein the concentrating of flag labeled microorganisms is on a flow strip device having one or more immobilization zones and the flag labeled microorganisms are captured and concentrated within the one or more immobilization zones.
Overall, the claim coverage is directed to a phage-based detection method that detects a flag on a microorganism bound via outside surface interaction without detecting amplified phage within the target, and that improves detection by concentrating the flag labeled microorganisms on immobilization zones of a flow strip device.
Stated Advantages
Enhances the capacity to detect the flag labeled microorganisms by concentrating the flag labeled microorganisms.
Avoids detecting phage amplified within the target microorganism.
Documented Applications
Detection of microorganisms using MS2 bacteriophage with lateral flow strips.
Detection of E. coli using the described approach.
Application of the approach to Salmonella, Bacillus anthracis, and group B Streptococcus using corresponding phages.
Interested in licensing this patent?