Mass Spectrometry Identification of EHEC Strains Within 6 Hours
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Solution Overview
Problem
Current methods are inadequate for rapid and accurate distinction between non-EHEC bacteria and specific EHEC strains associated with health risks, such as O157, O26, O45, 0103, 0104, O111, 0121, and/or 0145, which are responsible for severe illnesses like hemolytic uremic syndrome.
Innovation Solution
A method utilizing mass spectrometry, particularly liquid chromatography tandem mass spectrometry (LC-MS/MS), to analyze bacterial proteins by enzymatic digestion, generating peptides for comparison against reference data to identify EHEC strains based on unique proteomic profiles.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If traditional serotyping and genotyping methods are used to identify EHEC strains, then identification accuracy is maintained, but detection time and operational complexity increase significantly
Solution Approach 1:
The patent replaces traditional mechanical/biochemical serotyping methods with mass spectrometry-based proteomic analysis. The system uses MALDI-TOF mass spectrometry to generate spectral fingerprints of bacterial proteins, enabling rapid identification of EHEC strains within hours rather than days, while maintaining high accuracy through database comparison of spectral patterns.
Solution Approach 2:
The patent changes the detection parameter from genetic sequence analysis (traditional genotyping) to proteomic spectral fingerprinting. By analyzing the mass-to-charge ratio of protein ions and comparing spectral patterns against reference databases, the system achieves both speed and accuracy in EHEC strain identification without requiring time-consuming cultural or genetic manipulation steps.
2Adaptability or versatility
If comprehensive serotyping of multiple EHEC strains is performed, then detection coverage is improved, but method complexity and resource requirements increase
Solution Approach 1:
The patent creates a universal detection system using mass spectrometry that can simultaneously identify multiple EHEC serotypes (O157, O26, O45, O103, O104, O111, O121, O145) through a single analytical platform. The system uses a comprehensive reference database containing spectral fingerprints of various EHEC strains and non-EHEC E. coli, allowing one instrument and protocol to cover all major pathogenic serotypes without requiring separate testing procedures for each strain type.
3Productivity
If rapid detection methods are implemented, then detection speed is improved, but measurement accuracy may deteriorate
Solution Approach 1:
The patent creates digital copies of reference proteomic spectral fingerprints for various EHEC strains and stores them in a database. When analyzing unknown samples, the system compares the obtained mass spectrometry spectra against these reference copies using pattern recognition algorithms. This copying approach enables rapid automated identification while maintaining high accuracy, as the system matches spectral patterns rather than requiring time-consuming manual characterization.
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
Enables rapid and accurate identification of EHEC strains within 6 hours, distinguishing them from other E. coli serotypes and non-EHEC strains, without requiring prior genus or species identification.
Implementation Method 1
subjecting the sample or protein present therein to mass spectrometry to obtain mass spectrometry data
Implementation Method 2
liquid chromatography tandem mass spectrometry (LC-MS/MS)
Data Source
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AI summary
The invention relates to methods for detecting the presence or absence of an enterohemorrhagic Escherichia coli (EHEC) in a sample and/or identifying said EHEC. The invention further relates to such methods comprising subjecting the sample or protein therein to mass spectrometry to obtain mass spectrometry data, comparing the obtained mass spectrometry data with reference data for at least one EHEC strain with a specific serotype and optionally one or more non-EHEC strains and detecting the presence or absence of the EHEC.