Coated Rod Sample Transfer Tool for PCR Inhibitor Neutralization
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Solution Overview
Problem
Current direct PCR methods for whole blood and plant material face challenges such as PCR inhibitor presence, requiring complex sample preparation, high robustness of PCR systems, and difficulties in handling small blood volumes, which affect DNA amplification efficiency and gel electrophoresis analysis.
Innovation Solution
A rod-shaped sample transfer tool coated with active substances like heparin that inactivates PCR inhibitors, allowing for the direct transfer of small blood samples into PCR reactions, thereby improving DNA amplification and reducing the need for extensive sample preparation.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Ease of operation
If direct PCR method with minimal handling steps is used, then ease of operation is improved, but reliability deteriorates due to PCR inhibitors in blood
Solution Approach 1:
The transfer tool is pre-coated with active substances (heparin, dextran sulfate, or chaotropic salts) that inactivate PCR inhibitors. This preliminary preparation of the tool allows it to neutralize inhibitors upon contact with the sample during transfer, enabling direct PCR with minimal handling steps while maintaining reliability.
2Productivity
If blood volume is reduced to below 1 μl for PCR, then productivity is improved, but ease of operation deteriorates due to difficulty in handling submicroliter samples
Solution Approach 1:
The transfer tool automatically manages the submicroliter blood sample through its coated surface properties. The active substance coating facilitates direct contact and transfer of tiny blood volumes (below 1 μl) without requiring precise pipetting techniques, making submicroliter handling as easy as microliter handling while improving productivity through reduced PCR volume.
3Reliability
If lysis buffer and proteinase are added for sample preparation, then reliability is improved by inactivating PCR inhibitors, but device complexity increases
Solution Approach 1:
The inhibitor-inactivating function is extracted from the complex lysis buffer system and transferred to the simple transfer tool coating. Instead of adding multiple reagents (lysis buffer, proteinase, neutralization buffer) to the sample, the active substances are pre-applied to the tool, which then neutralizes inhibitors during the transfer process itself, eliminating the need for extensive sample preparation materials.
4Reliability
If incubation steps at elevated temperatures are performed, then reliability is improved by destroying PCR inhibitors, but loss of time increases
Solution Approach 1:
The incubation step for inhibitor destruction is skipped entirely. The active substance coating on the transfer tool rapidly inactivates PCR inhibitors during the transfer process itself, allowing the procedure to rush through to direct PCR without time-consuming incubation steps at elevated temperatures, thus saving 5-30 minutes while maintaining reliability.
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
The tool enables efficient DNA amplification in PCR reactions by binding or inactivating inhibitors, facilitating the use of small blood volumes and improving PCR robustness, while minimizing handling complexities and PCR system requirements.
Implementation Method 1
a rod-shaped or tubular sample transfer tool having at least one segment coated with one or more active substances, having an effect on a sample containing nucleic acids, so that materials contained in the sample that would inhibit an enzymatic reaction taking place after the transfer are bound at least in part, or inactivated
Data Source
AI summary
The invention relates to an active substance-coated rod-shaped sample transfer tool, the preparation thereof, and the use thereof for transferring nucleic acid-containing sample material into reaction mixtures of enzymatic detection reactions.