CRISPR-Cas LFA Test Strip Ratio Tuning to Reduce False Positives
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Solution Overview
Problem
CRISPR-Cas LFA tests often produce false positive results due to tolerance to variations in the amount of reporter molecule precursor introduced, leading to inaccurate nucleic acid detection.
Innovation Solution
The test kit is designed with an LFA test paper configuration where the ratio of labeled antibodies to capture molecules (A/S) is set to 1≤S/A, widening the allowable range for the amount of reporter molecule precursor introduced, thereby reducing false positives.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If the amount of reporter molecule precursor introduced is increased to improve detection sensitivity, then the detection sensitivity is improved, but false positive results increase due to tolerance variations
Solution Approach 1:
The patent changes the ratio parameter of labeled antibodies to capture molecules (A/S) to 1≤S/A, which widens the allowable range for reporter molecule precursor introduction and reduces false positives while maintaining detection sensitivity
Solution Approach 2:
The patent uses a composite configuration of labeled antibodies and capture molecules with specific ratio requirements, creating a robust test system that tolerates variations in precursor amounts
2Reliability
If the ratio of labeled antibodies to capture molecules is decreased to reduce false positives, then the robustness against precursor variation is improved, but the detection sensitivity may be reduced
Solution Approach 1:
The patent optimizes the ratio parameter S/A to be ≥1, which balances the competing requirements of robustness and sensitivity by widening the allowable precursor range while maintaining adequate signal generation
Solution Approach 2:
The patent creates a dynamic system where the labeled antibody and capture molecule amounts can vary within a broader range while maintaining the S/A ratio≥1, allowing flexibility in precursor amounts without losing detection capability
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 configuration enhances the robustness of the test by minimizing false positives and ensuring accurate detection of nucleic acids by maintaining the coloration of the first detection line under varying precursor amounts.
Implementation Method 1
a nucleic acid detection method (CRISPR-Cas LFA test) that combines CRISPR-Cas, which is a genome editing technique, and lateral flow assay (LFA), which is a test method based on an antigen-antibody reaction
Implementation Method 2
a nucleic acid detection method (CRISPR-Cas LFA test) that combines CRISPR-Cas, which is a genome editing technique
Implementation Method 3
a first detection line to which a capture molecule that captures the second antigen is fixed
Data Source
AI summary
A test kit for detecting a target nucleic acid sequence in a specimen according to an embodiment includes a lateral flow assay (LFA) test paper. The LFA test paper tests a reaction solution obtained by adding the specimen to a reaction solution containing a Cas enzyme and a reporter molecule precursor and subjecting the reaction solution to a CRISPR-Cas reaction. The LFA test paper includes a labeling area containing labeled antibodies, a first detection line to which capture molecules for capturing the reporter molecule precursor are fixed, and a second detection line to which capture antibodies for capturing the labeled antibodies are fixed. The amount A of the labeled antibodies and the amount S of the capture molecules on the LFA test paper satisfy conditions of A≥10−12 mol and 1≤S/A.