Microbial Cell Concentration via Selective Lysis and Cushioning Liquid
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Solution Overview
Problem
Current clinical microbiology methods are too slow for timely pathogen identification, leading to delayed antibiotic selection and increased resistance, as they rely on labor-intensive processes and require nucleic acid extraction and purification, which are time-consuming and often too late to impact patient outcomes in cases of sepsis or septic shock.
Innovation Solution
A method involving selective lysis of non-microbial cells followed by centrifugal separation and wash cycles to concentrate microbial cells, allowing for direct molecular assays without nucleic acid extraction, using a pretreatment vessel with a blood cell lysis reagent and a hydrophobic cushioning liquid to collect microbial cells, enabling rapid identification and antimicrobial de-escalation.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If standard microbial culture-based techniques are used for pathogen identification, then reliable pathogen identification can be achieved, but the time lag of 2-3 days prevents timely impact on patient outcomes
Solution Approach 1:
The patent applies preliminary action by performing selective lysis of host cells and centrifugal separation of microbial cells before nucleic acid extraction. This pre-treatment step concentrates microbial cells and removes inhibitors in advance, enabling faster downstream molecular analysis without compromising identification accuracy
Solution Approach 2:
The patent extracts and isolates microbial cells from the blood sample through selective lysis and centrifugal separation. By taking out the microbial cells and concentrating them at the interface, the method eliminates the need for time-consuming nucleic acid extraction and purification steps, reducing overall processing time while maintaining diagnostic accuracy
2Reliability
If nucleic acid extraction and purification steps are performed, then inhibitor-free samples are obtained for molecular assays, but the process takes more than 3 hours
Solution Approach 1:
The patent performs preliminary concentration and purification of microbial cells through selective lysis and centrifugal separation before molecular assays. This pre-treatment removes host cell debris and inhibitors in advance, providing clean samples for downstream applications without requiring time-consuming nucleic acid extraction and purification steps
Solution Approach 2:
The patent extracts microbial cells from the complex blood matrix through selective lysis of host cells and centrifugal separation. By taking out and concentrating the microbial cells at the liquid interface, the method obtains inhibitor-free samples directly, eliminating the need for prolonged nucleic acid extraction and purification processes
3Reliability
If broad-spectrum antibiotics are prescribed immediately for sepsis treatment, then patient survival is improved, but emergence of resistant strains increases
Solution Approach 1:
The patent implements feedback by providing rapid pathogen identification results that enable clinicians to reassess and adjust antibiotic therapy. The quick diagnostic information creates a feedback loop where treatment can be optimized based on actual pathogen identity, allowing de-escalation from broad-spectrum to targeted antibiotics while maintaining patient survival benefits
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 to result for nucleic acid assays, allowing for timely antibiotic selection and de-escalation from broad-spectrum to narrow-spectrum antibiotics, thereby reducing treatment costs and side effects, and improving patient outcomes by providing rapid microbial identification.
Implementation Method 1
selective lysis of non-microbial cells followed by centrifugal separation
Implementation Method 2
centrifugal separation and wash cycles to concentrate microbial cells
Implementation Method 3
a hydrophobic cushioning liquid to collect microbial cells
Data Source
Figure 1(a)~1(f)
Figure 1(g)~1(m)
Figure 2(a)~2(c)
AI summary
Methods and devices are provided for pretreatment of a sample containing microbial cells. In some embodiments, the pretreatment of the sample is performed via the initial selective lysis, within a sample pretreatment vessel, of non-microbial cells (such as blood cells) and the subsequent centrifugal separation of the sample to remove the resulting debris and concentrate the microbial cells. An immiscible and dense cushioning liquid may be included for collecting the microbial cells adjacent to the liquid interface formed by the cushioning liquid upon centrifugation of the pretreatment vessel. After removal of a substantial quantity of the supernatant, resuspension of the collected microbial cells, and re-establishment of the cushioning liquid interface, at least a portion of the remaining suspension may be removed without substantially removing the cushioning liquid. One or more intermediate wash cycles may be performed prior to extraction of the remaining suspension, which provides a "pretreated" sample.