Interferometric Bacterial Detection for Real-Time Phage Virulence
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Solution Overview
Problem
Current methods for quantifying phage virulence and measuring phage-host dynamics are labor-intensive, not amenable to automation, and limited to simple media, failing to capture phage-host interactions effectively.
Innovation Solution
Utilizing interferometry-based biosensors to detect and quantify phage virulence and phage-host dynamics in real-time, enabling label-free detection and analysis in complex media through interferometry systems that measure binding and lysis signals.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Extent of automation
If traditional microbiology double-layer agar assay is used to quantify phage virulence, then phage-host interactions can be detected, but the process is labor-intensive and not amenable to automation
Solution Approach 1:
The patent replaces the mechanical/manual operations of traditional microbiology assays (spotting phage suspensions, overnight cultures, manual plaque counting) with an optical measurement system based on interferometry. The system uses optical detection to monitor phage-host interactions in real-time, enabling automated high-throughput screening without manual intervention.
Solution Approach 2:
The patent implements continuous real-time monitoring of phage-host interactions using interferometric measurements. Instead of discrete endpoint measurements requiring overnight incubation, the system continuously tracks binding and lysis events as they occur, enabling automated decision-making and significantly improving throughput.
2Loss of time
If traditional microbiology double-layer agar assay is used, then phage virulence can be quantified, but it requires overnight cultures and is time-consuming
Solution Approach 1:
The patent performs preliminary functionalization of sensor surfaces with phage-specific capture molecules before sample introduction. This pre-preparation enables immediate real-time detection upon sample addition, eliminating the need for overnight cultures while maintaining accurate quantification of phage virulence through continuous monitoring.
Solution Approach 2:
The patent substitutes time-consuming manual plaque counting and overnight incubation with rapid optical interferometric detection. The system measures phage-host binding and lysis events in real-time, reducing detection time from overnight to minutes while maintaining measurement precision through sensitive optical detection.
3Loss of information
If traditional agar plate method is used, then phage-host interactions can be observed, but it cannot capture phage-host dynamics such as infectivity parameters
Solution Approach 1:
The patent replaces static endpoint observation on agar plates with dynamic real-time optical monitoring. The interferometric system continuously measures changes in mass or refractive index at the sensor surface, capturing phage-host binding kinetics, infection processes, and lysis events, thereby preserving complete phage-host dynamics information.
Solution Approach 2:
The patent implements continuous monitoring of phage-host interactions throughout the entire infection process. Instead of a single endpoint measurement, the system tracks binding events, infection progression, and lysis in real-time, capturing complete kinetic data and infectivity parameters without requiring complex additional instrumentation.
4Adaptability or versatility
If traditional microbiology assay is used, then phage infectivity can be measured, but it is limited to agar plates that do not ideally recapitulate infection conditions
Solution Approach 1:
The patent designs a universal sensor-based platform that can measure phage-host interactions in multiple media types including complex biological fluids. The interferometric detection method is not restricted to agar plates and can function in aqueous solutions, serum, and other physiologically relevant media, enhancing both adaptability and reliability.
Solution Approach 2:
The patent replaces the restrictive agar plate substrate with a versatile sensor surface that can be functionalized for different applications. The optical detection system works in various media without requiring solid support, enabling measurements in physiologically relevant conditions while maintaining measurement reliability.
Applied Scientific Principles
This section explains which scientific principles are used to turn an abstract innovation direction into a practical engineering solution.
Function Achieved in This Case
Enables high-throughput, automated, and label-free detection of phage-host interactions in complex media, providing real-time measurements of binding kinetics and lysis times, and identifying effective phages for personalized therapy.
Implementation Method 1
the general inventive concepts are based, in part, on the discovery that interferometry can be harnessed according to and along with the inventive concepts herein to detect the presence of pathogens, quantify phage virulence, and measure one or more phage-host dynamics and parameters
Implementation Method 2
irradiating the functionalized sensor with light and detecting and obtaining in real-time an at least one wavelength of white light reflected by the tip of the functionalized sensor with the bacteriophages attached thereto and corresponding to a baseline interference pattern measurement
Data Source
AI summary
Disclosed herein are systems and methods for quantifying bacteriophage virulence, measuring phage-host dynamics and parameters, including phage-host range, phage interactions with biological samples and with immune systems, and label-free bacterial detection/diagnostics, that are amenable to automation, high-throughput, and functional in complex media. In some embodiments, a label-free interferometry system transduces the light reflected by a sensor and any molecules attached thereto to a real-time signal comprising a sensorgram from which infectivity parameters such as binding kinetics and lysis time can be derived.