ELISPOT Assay for Mucorales-Specific T Cell Detection
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Solution Overview
Problem
Current diagnostic methods for invasive Mucorales infections are invasive, insensitive, and often delayed, leading to high mortality due to the lack of timely and certain diagnosis, as they rely heavily on histological and cultural examinations which are limited by invasiveness, contamination, and difficulty in isolating Mucorales from biological samples.
Innovation Solution
The use of immunoenzymatic assays, specifically the ELISPOT method, to identify and count Mucorales-specific T cells producing IFN-γ, IL-10, and IL-4 in biological fluids after exposure to Mucorales antigens, allowing for the detection and monitoring of active or previous infections, thereby improving clinical management.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If histological and cultural examinations are used for diagnosis, then diagnostic certainty can be achieved, but the diagnosis is delayed and mortality increases
Solution Approach 1:
The patent uses specific antibodies as intermediaries to detect Mucorales antigens directly in patient samples. These antibodies serve as mediators between the diagnostic system and the target pathogen, enabling rapid and certain diagnosis without requiring time-consuming cultural examinations or invasive histological procedures
Solution Approach 2:
The patent replaces the mechanical and time-intensive processes of histological examination and cultural cultivation with an immunoenzymatic assay system. This substitution uses biochemical reactions (enzyme-substrate interactions) to produce detectable signals, dramatically reducing diagnosis time while maintaining or improving diagnostic certainty
2Measurement precision
If biopsy samples are obtained for examination, then diagnostic accuracy improves, but patient invasiveness and difficulty increase
Solution Approach 1:
The patent extracts the essential diagnostic function from invasive biopsy procedures by detecting Mucorales-specific antigens directly in accessible body fluids such as serum or plasma. This extraction approach isolates the diagnostic capability from the invasive sampling requirement, maintaining accuracy while eliminating procedural difficulties
Solution Approach 2:
The patent employs specific antibodies as intermediaries that can detect Mucorales antigens in non-invasive or minimally invasive samples. These antibodies act as mediators that bridge the gap between easily obtainable biological fluids and the diagnostic detection system, eliminating the need for difficult biopsy procedures
3Measurement precision
If cultural examination is performed, then Mucorales detection is possible, but sensitivity remains low due to contamination and growth inhibition
Solution Approach 1:
The patent uses specific antibodies as intermediaries that selectively bind to Mucorales antigens with high affinity and specificity. This selective binding mechanism eliminates the problems of contamination and non-specific growth that plague cultural examinations, providing reliable and sensitive detection even in complex biological samples
Solution Approach 2:
The patent changes the detection parameter from observing fungal growth (which is slow and susceptible to contamination) to detecting specific antigen-antibody reactions (which are rapid and highly specific). This parameter change transforms the examination from a culture-based method with low sensitivity to an immunoenzymatic method with high 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
The ELISPOT method provides a sensitive and specific tool for diagnosing and monitoring invasive Mucormycosis, enabling early detection and improving patient outcomes by identifying specific T cell responses, thus addressing the limitations of existing diagnostic methods.
Implementation Method 1
The subject of the present invention is therefore a method for the diagnosis and/or monitoring of IM which comprises the execution of an immunoenzymatic or immunocytofluorimetric assay in vitro on microtitration plates
Implementation Method 2
immunoenzymatic assays (ELISPOT, Quantiferon) or immunocytofluorimetric assays [Cytokine Secretion Assay (CSA), Intracellular Cytokine Staining (ICS)]
Data Source
AI summary
A method is described for the diagnosis and/or monitoring of active or previous infection by Mucor which consists in the identification of Mucorales-specific T cells in samples from biological fluids taken from the patient and put into contact with a Mucor antigen. These specific immune responses can be detected by the execution of immunoenzymatic assays (ELISPOT, Quantiferon) or of immunocytofluorimetric assays [Cytokine Secretion Assay (CSA), Intracellular Cytokine Staining (ICS)] in vitro. In greater detail, the method in question provides for checking for the presence of specific IFN-γ producing T cells, of specific IL-10 producing T cells and/or specific IL-4 producing T cells.