Ricin Toxin Detection via Fluorescent Oligonucleotide Cleavage
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Solution Overview
Problem
Current ricin toxin detection methods are time-consuming, expensive, and have a narrow application range, limiting their effectiveness for rapid and sensitive detection.
Innovation Solution
A rapid detection method for ricin toxin involving the reaction of an adenine-containing oligonucleotide chain substrate labeled with fluorescent and quenching groups, along with a buffer solution, BSA solution, and a sample, at a constant temperature, followed by fluorescence signal detection and enrichment using antibody-coated magnetic beads.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If mass spectrometry technology is used for detection based on N-glycosidase activity, then detection sensitivity and specificity are improved, but detection time increases to 2-5 hours and instrument cost increases
Solution Approach 1:
The patent replaces mass spectrometry technology with fluorescence detection technology. The fluorescence-based detection system uses fluorescently labeled oligonucleotide substrates that emit fluorescence when cleaved by RT, enabling rapid detection within 30-60 minutes without requiring expensive mass spectrometry equipment, thus resolving the contradiction between detection sensitivity and detection time
Solution Approach 2:
The patent changes the detection parameter from mass spectrometry signal to fluorescence signal. By using fluorescently labeled substrates and measuring fluorescence intensity changes, the method achieves high sensitivity detection comparable to mass spectrometry but with significantly reduced detection time and equipment cost
2Measurement precision
If mass spectrometry technology is used for detection based on N-glycosidase activity, then detection sensitivity and specificity are improved, but instrument cost increases
Solution Approach 1:
The patent substitutes complex mass spectrometry instrumentation with a simple fluorescence detection system. The fluorescence detector is a standard, inexpensive instrument widely available in laboratories, eliminating the need for costly mass spectrometry equipment while maintaining detection sensitivity through fluorescent signal measurement
Solution Approach 2:
The patent uses disposable fluorescently labeled oligonucleotide substrates that are inexpensive to synthesize. These single-use substrates eliminate the need for expensive, reusable mass spectrometry instruments, providing a cost-effective detection solution with comparable sensitivity
3Adaptability or versatility
If conventional detection methods are used, then detection can be performed, but application scope is narrow and not suitable for on-site detection
Solution Approach 1:
The patent develops a universal fluorescence-based detection platform that can detect RT in various sample types including food, water, and environmental samples. The method uses standard reagents and equipment available in most laboratories, making it adaptable to different detection scenarios and suitable for on-site application
Solution Approach 2:
The patent employs a self-contained detection system where all necessary components (fluorescent substrates, buffer solutions, enrichment beads) are integrated into a single protocol. The method requires minimal external equipment and can be performed autonomously in various settings, enhancing its versatility and ease of on-site operation
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 method achieves high sensitivity and specificity, allowing for rapid detection of ricin toxin in as little as 40 minutes, with a sensitivity of 0.5 μg/mL without enrichment and 20 ng/mL with enrichment, and is suitable for on-site detection.
Implementation Method 1
an adenine-containing oligonucleotide chain substrate labeled with a fluorescent group and a quenching group
Implementation Method 2
detecting a fluorescence signal value
Implementation Method 3
enriching the to-be-detected sample by using antibody-coated magnetic beads
Implementation Method 4
antibody-coated magnetic beads
Implementation Method 5
RT is composed of two polypeptide chain subunits, A and B... The A chain mainly plays a toxic role... can specifically and irreversibly hydrolyze adenine (A4324) on the 28S rRNA of the ribosome
Data Source
AI summary
A rapid detection method for ricin toxin is provided. The rapid detection method for ricin toxin comprises reacting an adenine-containing oligonucleotide chain substrate labeled with a fluorescent group and a quenching group, a buffer solution, a BSA solution, a to-be-detected sample and sterile water, incubating at a constant temperature, detecting a fluorescence signal value, and determining whether the ricin toxin exists based on a difference between an average value of final fluorescence signal values and an average value of final fluorescence signal values of a negative control; wherein the sample is judged as a positive sample when the average value of the detected final fluorescence signal values is greater than the average value of the final fluorescence signal values of the negative control by +3 times standard deviation.