Dried Blood Spot Microsampling for Cystic Fibrosis Mutation Detection
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Solution Overview
Problem
Current methods for detecting cystic fibrosis-related mutations in biological samples are cumbersome, require large volumes of blood, and are not suitable for all patient groups, such as the elderly or infants, necessitating the development of rapid and non-invasive techniques for genetic diagnosis.
Innovation Solution
The use of a volumetric absorptive microsampling device to collect small-volume dried biological fluid samples, followed by extraction of CFTR nucleic acid and detection of mutations using high-throughput massive parallel sequencing, allowing for the identification of hereditary cystic fibrosis-related mutations in samples as small as 10-20 μL.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Quantity of substance
If conventional blood collection methods are used for detecting CFTR mutations, then sufficient nucleic acid can be obtained for analysis, but the procedure becomes cumbersome and requires large blood volumes that are not suitable for infants or elderly patients
Solution Approach 1:
The patent extracts only the necessary component (nucleic acid) from a small volume of dried blood spot, using targeted elution and extraction methods that recover sufficient CFTR gene material from just 10-20 μL of dried blood, eliminating the need for large blood volumes while maintaining analytical capability
Solution Approach 2:
The patent introduces a dried blood spot as an intermediary sample type between liquid blood and nucleic acid extraction. The dried blood spot stabilizes the nucleic acid while requiring minimal blood volume, and serves as a convenient intermediate that can be easily collected, stored, and processed without specialized equipment
2Reliability
If conventional nucleic acid extraction procedures are used, then complete genetic analysis can be performed, but the procedures become cumbersome and require specialized equipment or technical skill
Solution Approach 1:
The patent employs extraction buffers and protocols that automatically stabilize and release nucleic acids from the dried blood spot matrix without requiring complex instrumentation. The chemical composition of the elution buffer itself facilitates the extraction process, making the system self-sufficient and eliminating dependence on specialized equipment
Solution Approach 2:
The patent changes the physical state of the blood sample from liquid to dried form, which stabilizes the nucleic acid and simplifies handling. This parameter change (from liquid to solid/dried state) transforms a complex, equipment-intensive liquid handling process into a simple, equipment-free dried sample analysis workflow
3Quantity of substance
If large volumes of blood are collected for mutation detection, then adequate sample material is obtained, but the method becomes invasive and unsuitable for certain patient groups such as infants or elderly patients
Solution Approach 1:
The patent extracts maximum nucleic acid utility from minimal blood volume by using highly efficient elution and extraction protocols that recover sufficient CFTR gene material from just 10-20 μL of dried blood, eliminating the need for large invasive blood draws while maintaining diagnostic capability
Solution Approach 2:
The patent changes the sample collection approach from liquid blood requiring large volumes to dried blood spots requiring minimal volumes. This parameter change (volume reduction through drying) transforms an invasive large-volume collection into a minimally invasive small-volume collection that is suitable for fragile patient populations
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 enables efficient and non-invasive detection of cystic fibrosis mutations in small samples, facilitating the selection of appropriate therapeutic agents for patients and carriers, while overcoming the limitations of existing techniques in terms of sample volume and patient suitability.
Implementation Method 1
volumetric absorptive microsampling device
Implementation Method 2
absorbent tip of a microsampling device
Implementation Method 3
contacting the absorbent tip of the microsampling device with a lysis buffer and Proteinase K
Implementation Method 4
elution of the dried biological fluid sample is performed by contacting the absorbent tip of the microsampling device with a lysis buffer
Data Source
AI summary
The present disclosure provides methods for determining whether a patient exhibiting cystic fibrosis symptoms, or a patient at risk for cystic fibrosis, will benefit from treatment with one or more anti-cystic fibrosis therapeutic agents. These methods are based on detecting hereditary cystic fibrosis related mutations in small-volume dried biological fluid samples that are collected using a volumetric absorptive microsampling device. Kits for use in practicing the methods are also provided.