Blood Sample Pooling via Lysis and Anticoagulation
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Solution Overview
Problem
Current blood screening methods for transfusions are inadequate for detecting pathogens like Babesia microti, as they cannot efficiently handle whole blood samples due to clotting issues and are limited in detecting pathogens within red blood cells, leading to potential transfusion-transmitted diseases.
Innovation Solution
A method involving a lysis reagent with lithium lauryl sulfate, a buffer, and an anticoagulant is used to lyse blood cells, allowing for pooling and subsequent nucleic acid testing to detect pathogens, including Babesia microti, by releasing target molecules for analysis in a pooled lysate.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Productivity
If whole blood samples are pooled directly for screening, then the proportion of infected samples can be reduced and screening simplified, but clotting occurs when samples from different patients are mixed which interferes with subsequent assay steps
Solution Approach 1:
The patent applies preliminary action by adding anticoagulant to each individual blood sample before pooling. This prevents clotting from occurring during the pooling process and subsequent assay steps. The anticoagulant is incorporated in advance into the collection tubes, ensuring that when samples are combined, they remain liquid and suitable for processing without forming clots that would interfere with the assay.
Solution Approach 2:
The patent extracts the harmful clotting factor by introducing anticoagulant that binds calcium ions, removing the ability of blood to clot. The anticoagulant selectively targets and neutralizes the clotting mechanism, allowing the blood samples to be pooled and processed without the formation of interfering clots while preserving other blood components for pathogen detection.
2Reliability
If plasma or serum is used for nucleic acid testing, then virus screening can be performed, but plasma and serum cannot be used to detect pathogens of red blood cells
Solution Approach 1:
The patent applies universality by using whole blood as a single sample type that can detect multiple types of pathogens - both viruses in plasma/serum and intracellular pathogens in red blood cells. By maintaining the whole blood matrix and preventing clotting with anticoagulant, the system achieves multi-functionality, allowing a single testing approach to screen for diverse pathogens including Babesia microti, HIV, hepatitis viruses, and others without requiring separate plasma or serum preparations.
3Measurement precision
If individual blood samples are tested separately, then accurate detection of infected samples is achieved, but the screening process becomes complex and time-consuming
Solution Approach 1:
The patent applies merging by combining multiple individual blood samples into a single pooled sample for simultaneous analysis. By pooling anticoagulated whole blood samples and performing nucleic acid testing on the combined sample, the method maintains the ability to detect infected samples while significantly reducing the number of separate tests required. If the pool tests positive, deconvolution procedures can identify the specific infected donor, but the initial screening is streamlined through pooling.
Solution Approach 2:
The patent applies self-service through the use of internal controls and automated deconvolution procedures. The pooled sample testing approach with built-in controls allows the system to self-identify positive samples, and when needed, automatically guide the deconvolution process to pinpoint infected donors without requiring extensive manual intervention or complex individual testing protocols for every sample.
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 effectively detects pathogen presence in whole blood samples, enabling the identification of infected samples within a pool, ensuring safer blood transfusions by discarding infected units and retaining uninfected ones for use.
Implementation Method 1
separately performing a lysing reaction on each of the sample aliquots, wherein the lysing step comprises contacting each of the sample aliquots with a lysis reagent comprising: a buffer, lithium lauryl sulfate (LLS), and at least one of a chloride containing salt and an anticoagulant
Implementation Method 2
an anticoagulant selected from the group consisting of: EDTA, EDTA-Na2, EGTA, and combinations thereof
Data Source
AI summary
A method for detecting the presence of a pathogen in a whole blood sample aliquot, includes providing a sample aliquot from each of a number of whole blood samples, thereby providing a plurality of whole blood sample aliquots, separately contacting each of the whole blood sample aliquots of the plurality of whole blood sample aliquots with a lysis reagent, whereby at least a portion of the blood cells in each of the whole blood sample aliquots lyse, pooling the lysed whole blood sample aliquots to form a pooled lysate, and testing the pooled lysate for presence of a pathogen. Identification of the presence of the pathogen in the pooled lysate indicates the presence of the pathogen in at least one of the whole blood sample aliquots.