Multiplex PCR Assay for Tick-Borne Co-Infection Detection
Find Innovative SolutionsGenerate Solutions
Solution Overview
Problem
Current diagnostic tests for Lyme disease and other tick-borne illnesses, such as anaplasmosis and babesiosis, are inadequate as they cannot accurately detect early infections, distinguish between active and cured infections, and fail to detect co-infections, leading to a need for a technically simple, rapid, and accurate assay to diagnose these diseases individually or simultaneously.
Innovation Solution
The development of multiplex primer-dependent assays, including PCR methods, that utilize molecular beacon probes to detect specific genetic targets of Borrelia burgdorferi, Babesia microti, and Anaplasma phagocytophilum, allowing for real-time amplification and differentiation of these pathogens in a single assay, with the option to include a human DNA target for control and quality assessment.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If serological tests are used to diagnose tick-borne diseases, then the tests can detect infections, but they cannot detect early infections before antibodies are produced and cannot distinguish between active and cured infections
Solution Approach 1:
The patent replaces serological testing (mechanical/chemical detection of antibodies) with nucleic acid-based detection (molecular-level detection of pathogen DNA/RNA). This substitution enables detection of the actual pathogen rather than the host's immune response, allowing for early detection before antibodies are produced and accurate differentiation between active and cured infections by detecting pathogen genetic material presence or absence.
2Measurement precision
If current nucleic acid-based assays are used, then they can specifically detect the presence of pathogens, but they are insufficiently multiplex to accurately detect co-infections
Solution Approach 1:
The patent merges multiple nucleic acid detection assays into a single multiplexed platform that can simultaneously detect and differentiate multiple tick-borne pathogens (Borrelia burgdorferi, Anaplasma phagocytophilum, Babesia microti) in one test. This combining of detection capabilities maintains the specificity of nucleic acid-based detection while adding the versatility to handle co-infections, resolving the contradiction between precision and adaptability.
3Measurement precision
If individual tests are developed for each pathogen, then accurate diagnosis of each disease is possible, but the complexity and time required for diagnosis increases
Solution Approach 1:
The patent creates a universal nucleic acid detection platform that can test for multiple tick-borne pathogens simultaneously through multiplexing. This single multi-functional assay replaces the need for separate individual tests for each pathogen, maintaining diagnostic accuracy for each disease while reducing overall system complexity and streamlining the diagnostic workflow.
4Ease of operation
If current diagnostic assays are used, then they can provide diagnosis, but they cannot provide accurate information about whether pathogens were introduced by ticks during bloodmeal
Solution Approach 1:
The patent replaces indirect serological detection with direct nucleic acid detection of the pathogen itself, which provides information about active infection status and can indicate recent transmission. By detecting pathogen genetic material rather than host antibodies, the system preserves information about whether the infection is currently active and likely recent, enabling more accurate determination of infection source and timing.
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
This approach enables sensitive and specific detection of active tick-borne infections, allowing for accurate diagnosis and differentiation of multiple pathogens simultaneously, improving the ability to diagnose co-infections and determine appropriate treatment regimens.
Implementation Method 1
a primer pair and a molecular beacon probe for each intended target, wherein the primer pair defines an amplification product ('amplicon') that is 70-300 base pairs in length, and wherein each different molecular beacon probe is labeled with a spectrally distinguishable fluorescent or luminescent signaling moiety
Implementation Method 2
each different molecular beacon probe is labeled with a spectrally distinguishable fluorescent or luminescent signaling moiety
Implementation Method 3
Nucleic acid-based assays, on the other hand, are able to specifically detect the presence of the pathogens that cause Lyme disease and other tick-borne diseases
Data Source
AI summary
The present invention provides novel methods of diagnosing and determining treatment strategies for Lyme disease and other tick-borne illnesses.