Food Sample Bacterial Concentration via Protease Pre-treatment and Filtration
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Solution Overview
Problem
Current methods for detecting bacteria in food samples are inefficient due to the complexity and diversity of food matrices, requiring significant enrichment time and large sample volumes, leading to poor representativeness and low yield, especially when contamination levels are low.
Innovation Solution
A method involving a two-step filtration process using proteases to degrade proteins and improve filtration, allowing for the concentration and purification of target bacteria, using filters with specific pore sizes and a preparation device for efficient sample preparation, enabling quick analysis with high bacterial concentration from a limited initial sample.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If traditional detection methods are used, then bacteria can be detected, but the process requires significant enrichment time (7-26 hours) and large sample volumes
Solution Approach 1:
The patent applies preliminary action by performing protease treatment and filtration steps before the actual bacterial detection. The method pre-processes the food matrix by degrading proteins that interfere with detection, concentrating bacteria through filtration, and removing inhibitors beforehand. This preliminary preparation eliminates the need for long enrichment periods (7-26 hours) while maintaining detection sensitivity, reducing the process to just a few hours.
2Measurement precision
If large sample volumes are used to obtain sufficient bacteria, then detection sensitivity improves, but the representativeness of the sample decreases
Solution Approach 1:
The patent applies the extraction principle by selectively separating target bacteria from the complex food matrix through filtration. The method extracts bacteria from large volumes of food matrix (up to 100 mL or more) by passing them through filters with specific pore sizes, concentrating the bacteria in a small final volume while maintaining the representativeness of the original sample. This allows achieving detection sensitivity equivalent to large sample volumes using only small final sample quantities.
3Quantity of substance
If filtration is performed on complex food matrices, then bacteria concentration improves, but filter clogging occurs due to proteins and debris
Solution Approach 1:
The patent applies preliminary action by treating the food matrix with proteases before filtration to degrade proteins and debris that would otherwise clog the filters. This pre-treatment step simplifies the filtration process by removing interfering substances, allowing direct filtration of complex food matrices without filter clogging, and enabling efficient bacterial concentration.
4Loss of time
If rapid detection techniques are used, then analysis time is reduced, but sensitivity and bacterial concentration remain insufficient
Solution Approach 1:
The patent applies the merging principle by combining multiple functions into a single integrated filtration device. The device simultaneously concentrates bacteria, removes interfering substances, and prepares samples for detection in one operation. This merged approach achieves both rapid processing (reducing time losses) and high sensitivity (through effective concentration and purification), eliminating the trade-off between speed and 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 enhances filtration efficiency, reduces sample volume requirements, and improves bacterial recovery, providing a more representative and efficient analysis of bacteria in food samples, compatible with techniques like q-PCR, LAMP, and sequencing.
Implementation Method 1
a second dilution of the sample already diluted during the previous step with a liquid containing a protease
Implementation Method 2
This filtration step is implemented by using a second filter F2 having pores of a size smaller than that of the pores of the first filter F1 used during prefiltration
Data Source
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AI summary
The invention relates to a method for preparing a sample from a food matrix sample, said sample containing biological species to be analyzed, the method comprising: - A step (E3) of diluting a first sample from said sample with a protease for a sufficient time to degrade said first sample and obtain a second liquid sample, - A step (E4) of pre-filtering said second liquid sample through a first filter (F1) to separate first molecules (M1) which are larger than the pores of said first filter (F1), in order to obtain a third sample comprising the biological species to be analyzed and second molecules (M2) smaller than the pores of said first filter, - A step (E5) of filtering said third sample through a second filter (F2) to separate said biological species from said second molecules (M2).