RT-qPCR T Cell Detection via APC Marker Induction
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Solution Overview
Problem
Current methods for detecting activated T cell populations, particularly antigen-specific CD4+ T cells, are limited in sensitivity and specificity, making it difficult to accurately diagnose active infections and autoimmune diseases due to the transient nature and low numbers of these cells in patient samples.
Innovation Solution
A method involving reverse transcription quantitative real-time polymerase chain reaction (RT-qPCR) is used to detect and quantify T cell populations by measuring markers induced in antigen-presenting cells (APCs) after interaction with activated T cells, allowing for the differentiation of naive, activated, and memory T cells through the detection of mRNA markers.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If current methods are used to detect activated T cell populations, then the detection process is simple, but the sensitivity and specificity are limited
Solution Approach 1:
The patent uses antigen-presenting cells (APCs) as an intermediary to detect activated T cells. Instead of directly detecting T cells, the method detects markers induced in APCs after they interact with activated T cells. This intermediary approach amplifies the signal and enables detection of transient, low-number T cell populations with high sensitivity and specificity using RT-qPCR technology.
Solution Approach 2:
The patent replaces traditional flow cytometry or immunohistochemistry methods with molecular biology-based RT-qPCR detection. By substituting mechanical/optical detection systems with biochemical amplification and detection systems, the method achieves superior sensitivity and specificity for detecting activated T cell populations.
2Reliability
If direct detection of T cells is performed, then the procedure is straightforward, but the reliability is low due to transient nature and low numbers of activated T cells
Solution Approach 1:
The patent performs preliminary incubation of APCs with antigens before detection to ensure activated T cells are present and have induced markers in APCs. This preliminary action increases the reliability of detection by ensuring the target signal is amplified and stable before the actual RT-qPCR measurement occurs.
Solution Approach 2:
By using APCs as mediators that accumulate and amplify signals from activated T cells, the method overcomes the reliability issues associated with directly detecting transient, low-number T cell populations. The intermediary APCs serve as signal amplifiers that make the detection more reliable.
3Measurement precision
If different T cell populations are to be differentiated, then specific marker detection is required, but the complexity of detection increases
Solution Approach 1:
The patent differentiates T cell populations by detecting specific markers in APCs that are induced by different T cell subsets. Each T cell population (e.g., Th1, Th2, Th17) induces characteristic markers in APCs, allowing segmentation and differentiation of T cell populations through targeted marker detection using RT-qPCR with specific primers for each marker.
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 enhances sensitivity and reliability in detecting activated T cells, enabling better discrimination between different T cell populations and aiding in the differential diagnosis of infectious and autoimmune diseases.
Implementation Method 1
detection and differentiation of the T cell populations by detecting at least a first marker of the APCs induced by the T cells of a specific T cell population in the first aliquot and in a second aliquot of the body fluid of the individual, which has not been incubated with the at least one antigen, by way of reverse transcription quantitative real time polymerase chain reaction (RT-qPCR)
Implementation Method 2
reverse transcription quantitative real time polymerase chain reaction (RT-qPCR)
Data Source
AI summary
The present invention relates to a method for detection, differentiation and quantification of T cell populations, comprising the following steps a) contacting a first aliquot of a body fluid of an individual with at least one antigen, wherein the body fluid contains antigen presenting cells (APC) and T cells, b) incubating the first aliquot with at least one antigen for a certain period of time, c) detection and differentiation of the T cell population by detecting in the first aliquot and in a second aliquot of the body fluid of the individual, which has not been incubated with the at least one antigen, at least a first marker of the APC induced by T cells in a specific T cell population using reverse transcription quantitative real time-time polymerase chain reaction (RT-qPCR), and d) detection and quantification of the T cell population by determining the ratio of the detected marker of the APC of the first aliquot to the second aliquot as well as a kit for performing the method.