DNA Aptamer for CFP10 Tuberculosis Biosensor
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Solution Overview
Problem
Current diagnostic methods for latent tuberculosis, such as tuberculin skin test and interferon-gamma releasing assay, face challenges including long duration, false-positive results, and reduced sensitivity, particularly in hot or humid environments, necessitating a more accurate and stable detection method.
Innovation Solution
A DNA aptamer specifically binding to culture filtrate protein 10 kDa (CFP10) is developed, comprising a nucleotide sequence, which is used in a biosensor to diagnose tuberculosis by immobilizing the aptamer on a metal electrode layer, preferably gold, and measuring the bound protein levels in subject samples.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If tuberculin skin test is used for latent tuberculosis diagnosis, then diagnostic coverage is achieved, but false-positive results occur due to BCG vaccination or non-tuberculous mycobacteria infection
Solution Approach 1:
The invention extracts and targets only the specific CFP10 antigen from Mycobacterium tuberculosis, excluding other mycobacterial antigens that cause false positives in traditional tests. The aptamer-based biosensor specifically binds to CFP10, eliminating cross-reactivity with BCG vaccine strains or non-tuberculous mycobacteria.
Solution Approach 2:
The invention uses a highly specific DNA aptamer that recognizes only the CFP10 epitope with precise local binding characteristics. This localized specific binding at the aptamer-CFP10 interface ensures high diagnostic accuracy without cross-reactivity, achieving both sensitivity and specificity simultaneously.
2Reliability
If interferon-gamma releasing assay is used for latent tuberculosis diagnosis, then specific antigen detection is achieved, but sensitivity is reduced due to indirect measurement of IFN-g secretion
Solution Approach 1:
Instead of indirectly measuring IFN-g secretion, the invention directly detects CFP10 antigen using an aptamer that serves as a molecular copy or surrogate for antibody-based detection. This direct antigen-aptamer binding provides both high specificity and enhanced sensitivity compared to the indirect ELISA method.
Solution Approach 2:
The invention replaces the complex cellular immune response measurement (IFN-g secretion requiring cell stimulation and supernatant analysis) with a direct molecular binding assay. The aptamer-CFP10 interaction provides immediate, quantitative detection without requiring biological sample processing or indirect measurement steps.
3Reliability
If interferon-gamma releasing assay is used for latent tuberculosis diagnosis, then specific antigen detection is achieved, but stability is reduced in hot or humid environments
Solution Approach 1:
The invention uses a stable DNA aptamer molecule that is chemically robust and resistant to degradation in hot or humid conditions. Unlike protein-based antibodies or cellular assays, the nucleic acid-based aptamer maintains structural integrity and binding affinity across varying environmental conditions, providing assay stability where traditional methods fail.
4Measurement precision
If traditional antibody-based detection methods are used, then high affinity for target is achieved, but false-negative results occur and stability is reduced
Solution Approach 1:
The invention changes the molecular class from protein-based antibodies to nucleic acid-based aptamers. This parameter change provides several advantages: aptamers can be engineered with optimized binding kinetics, exhibit higher thermal and chemical stability, and avoid batch-to-batch variability inherent in antibody preparations, thereby reducing false-negative results.
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 DNA aptamer provides high specificity and stability, excluding false-negative and false-positive responses, offering a reliable diagnostic tool for tuberculosis with improved sensitivity and stability compared to existing methods.
Implementation Method 1
A DNA aptamer specifically binding to culture filtrate protein 10 kDa (CFP10) is developed
Data Source
AI summary
The present invention relates to: a DNA aptamer specifically binding to culture filtrate 10 kDa (CFP10); a biosensor for diagnosing tuberculosis, comprising the same: and an information providing method for diagnosing tuberculosis. The present applicants have ascertained that the DNA aptamer, according to the present invention, has the ability to specifically bind to a CFP10 protein and that the binding strength thereof is strong. When the DNA aptamer of the present invention is used, excellent stability superior to that of ELISA methods using existing antibodies can be expected, and thus it is expected that the aptamer can be effectively used in the development of a composition for diagnosing tuberculosis, a biosensor for diagnosing tuberculosis, an information providing method for diagnosing tuberculosis, and the like.