STEC Detection Agar with Nanoparticle Halo Formation
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Solution Overview
Problem
Current methods for detecting and confirming virulent Escherichia coli producing Shiga toxin (STEC) are time-consuming and prone to false negatives, especially in polymicrobial samples, due to the need for multiple amplification and confirmation steps.
Innovation Solution
A method involving sample lysis, nucleic acid amplification of stx1 and stx2 genes, and deposition on an agar reaction medium with toxin inducers and specific binding partners coupled to nanoparticles, allowing for direct visualization of STEC presence through halo formation.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If multiple amplification and confirmation steps are used to detect STEC, then detection reliability is improved, but detection time increases and productivity decreases
Solution Approach 1:
The patent combines multiple detection steps (amplification, immunoconcentration, and confirmation) into a single integrated assay performed on a single Petri dish. The agglutination test is conducted directly on the dish containing amplified bacterial colonies, eliminating the need for separate confirmation steps and enabling simultaneous detection and visualization of STEC.
Solution Approach 2:
The Petri dish serves multiple functions: it acts as both the growth medium for bacterial amplification and the substrate for the agglutination confirmation test. This multi-functional approach allows the same platform to perform both detection and confirmation roles, streamlining the overall process.
2Reliability
If multiple amplification and confirmation steps are used to detect STEC, then detection reliability is improved, but device complexity increases
Solution Approach 1:
The patent merges the amplification and confirmation procedures into a single integrated assay performed on one Petri dish, reducing the number of separate devices and操作步骤 required. The agglutination test is executed directly on the dish containing the amplified colonies, simplifying the overall method.
3Reliability
If traditional detection methods are used in polymicrobial samples, then comprehensive detection is achieved, but false negatives increase due to difficulty in locating target microorganisms
Solution Approach 1:
The patent utilizes color changes through the agglutination reaction to visually identify STEC colonies among polymicrobial samples. The formation of visible agglutinates or color changes on the Petri dish provides a clear visual signal that distinguishes target STEC colonies from non-target organisms, significantly improving target identification accuracy.
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 method enables rapid and accurate detection and confirmation of STEC, reducing the risk of false negatives and streamlining microbiological control by visually identifying target microorganisms through distinct halo formations.
Implementation Method 1
at least one agglutinating conjugate formed by at least one specific binding partner of the STX1 protein and/or at least one specific binding partner of the STX2 protein, coupled to a nanoparticle
Data Source
Figure 1~2
AI summary
The invention relates to a method for the detection and confirmation of at least one Shiga toxin-producing Escherichia coli (STEC) likely to be present in a sample containing enterobacteria, comprising the following steps: - Performing lysis of the sample enabling the lysis of STEC to obtain a solution comprising their nucleic acids - Contacting the nucleic acid solution with primers enabling the amplification of at least the stx1 and/or stx2 gene or gene fragment - If at least one of the stx1 and/or stx2 genes or gene fragments is amplified, a portion of the sample is deposited on an agar reaction medium comprising ■ at least one toxin inducer, ■ at least one agglutinating conjugate formed by at least one specific binding partner of the STX1 protein and/or at least one specific binding partner of the STX2 protein, coupled(s) to a nanoparticle.- Detect and confirm the presence of at least one STEC by the appearance of a halo on the agar around said STEC.