Protease Buffer Microbial Cell Recovery Filtration
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Solution Overview
Problem
Current methods for diagnosing microbial infections, particularly in cases of sepsis, are hindered by the need for prolonged incubation periods to isolate and culture microorganisms, leading to delayed antimicrobial susceptibility testing and increased use of broad-spectrum antibiotics, which fuels antimicrobial resistance.
Innovation Solution
A method involving the use of buffer solutions containing proteases to enhance the filterability of complex samples, allowing for the rapid recovery of viable microbial cells without the need for complete lysis of non-microbial cells, thereby enabling efficient microbial recovery and susceptibility testing from larger sample volumes.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If conventional culture methods are used to isolate microorganisms, then microbial identification and susceptibility testing can be performed, but the process requires prolonged incubation periods of several days
Solution Approach 1:
The patent applies preliminary action by performing protease treatment on the clinical sample before filtration to pre-digest proteins and reduce sample complexity. This preparatory step enables faster subsequent processing and reduces the overall time required for microbial identification while maintaining accuracy
Solution Approach 2:
The patent replaces the traditional mechanical/physical culture-based identification system with a biochemical approach using protease enzymes to digest proteins and release microbial components. This substitution allows for rapid detection methods rather than relying on prolonged incubation and visual observation of growth
2Reliability
If broad-spectrum antibiotics are used initially to treat sepsis, then patient survival is improved, but antimicrobial resistance increases
Solution Approach 1:
The patent implements feedback by rapidly identifying the specific microorganism causing infection through protease-based digestion and detection methods. This quick identification provides feedback to clinicians, allowing them to adjust treatment from broad-spectrum antibiotics to targeted therapy, thereby reducing antimicrobial resistance while maintaining patient survival rates
3Productivity
If complete lysis of non-microbial cells is performed to recover microbial cells, then microbial recovery efficiency is improved, but the complexity of the procedure increases
Solution Approach 1:
The patent applies the extraction principle by using proteases to specifically digest and break down proteins in the complex clinical sample matrix. This selectively removes interfering proteins and releases microbial cells without requiring complete lysis of non-microbial cells, simplifying the procedure while maintaining high recovery efficiency
Solution Approach 2:
The patent changes the chemical parameters of the sample by adding proteases that alter the protein structure and solubility. This parameter change enables selective digestion of proteins while preserving microbial cell integrity, achieving high recovery efficiency without complex lysis procedures
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 approach significantly reduces the time required for microbial identification and susceptibility testing, potentially limiting the use of broad-spectrum antibiotics and mitigating antimicrobial resistance by enabling rapid and effective treatment.
Implementation Method 1
contacting said sample with a buffer solution and one or more proteases
Data Source
AI summary
The present invention provides a method of recovering viable microbial cells from a complex sample, said method comprising: a) providing a sample having a volume of at least 1 ml; b) contacting said sample with a buffer solution and one or more proteases, wherein said buffer solution has a pH of at least pH 6 and less than pH 11, wherein said buffer solution and said one more proteases do not comprise a detergent or a chaotrope, and wherein the buffer solution/protease/sample mixture is non-hypotonic; c) filtering the mixture obtained in step (b) through a filter suitable for retaining microbial cells; and d) recovering the microbial cells retained by the filter in step (c), wherein the recovered microbial cells are viable, and a microbial recovery device for the same.