Histamine Aptamer Enzyme Sensor for Aquatic Freshness Detection
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Solution Overview
Problem
Current nucleic acid aptamers for histamine exhibit insufficient affinity, necessitating multiple iterative optimizations, and there is a need for a more efficient method to enhance their detection specificity and sensitivity in aquatic product freshness evaluation.
Innovation Solution
A high-affinity nucleic acid aptamer (HIS3-T2) with a modified sequence (SEQ ID NO. 2) is developed, combined with an aquatic product freshness enzyme sensor comprising chitopentaose, nanozyme AuNPs@FeP, and a TMB chromogenic solution, enabling rapid and specific detection of histamine.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If truncation-based optimization is applied to enhance aptamer affinity, then the affinity for histamine detection is improved, but the optimization process requires multiple attempts and increased complexity
Solution Approach 1:
The patent applies preliminary action by pre-modifying the aptamer sequence with specific nucleotide changes (G to A substitutions at positions 13, 23, and 43) before the detection process. This pre-optimization of the aptamer sequence structure enables high affinity (Kd = 59.48 nM) to be achieved in a single design iteration, eliminating the need for multiple truncation attempts and reducing optimization complexity.
2Measurement precision
If conventional aptamer detection methods are used, then the detection capability is achieved, but the detection sensitivity and specificity are insufficient
Solution Approach 1:
The patent applies parameter changes by modifying the aptamer's nucleotide sequence parameters (specific G to A substitutions) to optimize binding characteristics. Additionally, the detection system uses colorimetric parameter changes (absorbance measurements at 650 nm) to achieve high detection sensitivity and specificity for histamine in aquatic products.
Solution Approach 2:
The patent employs composite materials by combining the modified aptamer HIS3-T2 with gold nanoparticles (AuNPs) to create a hybrid detection system. This composite structure enhances both the sensitivity and specificity of histamine detection, allowing for reliable differentiation of histamine from other biogenic amines through specific aptamer-AuNP interactions.
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 aptamer HIS3-T2 achieves a 25.97-fold increase in affinity for histamine, allowing for rapid and accurate freshness evaluation of aquatic products by detecting histamine concentrations using microneedle patches and an enzyme sensor.
Implementation Method 1
Nucleic acid aptamers are a class of single-stranded nucleotides with specific recognition capabilities, which can achieve high-specificity and high-affinity binding to targets
Implementation Method 2
an aquatic product freshness enzyme sensor comprising chitopentaose, nanozyme AuNPs@FeP, and a TMB chromogenic solution
Implementation Method 3
TMB chromogenic solution, where in the chromogenic solution, the TMB has a concentration of 2 mM
Implementation Method 4
using microneedle patches to perform pressing extraction on an aquatic product to be detected for 2 min
Data Source
AI summary
A high-affinity nucleic acid aptamer for histamine and an aquatic product freshness enzyme sensor, belonging to the technical field of biological detection, is named HIS3-T2 and has a nucleotide sequence shown in SEQ ID NO.2. An application of the high-affinity nucleic acid aptamer for histamine in detecting, separating or enriching the histamine. The aquatic product freshness enzyme sensor includes the following components: an aptamer HIS3-T2, chitopentaose, a nanozyme AuNPs@FeP, and a TMB chromogenic solution. Applications of the aquatic product freshness enzyme sensor in detecting the histamine and in rapidly evaluating freshness of an aquatic product. The present invention further provides a method for rapidly evaluating freshness of an aquatic product. The present invention provides a new biological recognition element for detection of the histamine, which has great significance for the detection and enrichment of the histamine.