Internal Control Biomolecule for Nucleic Acid Detection
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Solution Overview
Problem
Current nucleic acid detection methods face challenges in minimizing false results due to inhibitory substances and reaction conditions, particularly when using amplification techniques, which can lead to misleading results and require the use of control nucleic acids to verify the accuracy of the detection process.
Innovation Solution
The method involves adding an internal control biomolecule to the test sample and control samples, allowing for the verification of the presence or absence of a target biomolecule by comparing signals across different samples, including a positive control, negative control, and reagent control, to ensure the reliability of the detection results.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If amplification methods are used to detect low concentration target nucleic acids, then detection sensitivity is improved, but the risk of false results due to inhibitory substances and reaction condition variations increases
Solution Approach 1:
An internal control nucleic acid is introduced as an intermediary element in the amplification reaction. This control nucleic acid undergoes amplification under the same conditions as the target nucleic acid but produces a distinguishable signal. The control serves as a mediator to monitor reaction suitability and detect inhibitory substances, allowing assessment of whether the amplification process is functioning correctly without directly affecting the target detection.
Solution Approach 2:
The internal control nucleic acid provides feedback information about the amplification reaction status. By comparing the control signal with the target signal, the system can determine whether the amplification process is proceeding normally or is compromised by inhibitors. This feedback mechanism enables real-time assessment of reaction conditions and allows for identification of false positives or false negatives.
2Reliability
If control nucleic acids are added to monitor assay performance, then reliability of detection is improved, but device complexity and procedure steps increase
Solution Approach 1:
The internal control nucleic acid is merged with the target nucleic acid detection process. Both the target and control nucleic acids are amplified in the same reaction chamber under identical conditions, eliminating the need for separate control reactions. The control nucleic acid is co-processed with the sample, reducing the number of separate steps and reagent additions while maintaining reliability monitoring.
Solution Approach 2:
The internal control nucleic acid serves multiple functions simultaneously: it acts as a positive control to verify amplification capability, a negative control to detect inhibitory substances, and a reference for quantification. This multi-functionality reduces the need for multiple separate control experiments, thereby reducing overall procedure complexity while maintaining comprehensive reliability checking.
3Reliability
If internal control nucleic acid is co-amplified with target nucleic acid in the same reaction chamber, then reliability monitoring is improved, but signal differentiation and assay complexity increase
Solution Approach 1:
The internal control nucleic acid is designed with local quality differences that enable signal differentiation. This includes using nucleic acids with distinct sequence characteristics, lengths, or structural features that produce distinguishable signals during detection. The control nucleic acid may have modified properties such as different probe binding characteristics or signal emission properties that allow clear differentiation from the target signal while maintaining co-amplification.
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 enhances the accuracy of nucleic acid detection by allowing for the monitoring of assay performance and the identification of potential inhibitors, thereby reducing false results and improving the reliability of the detection process.
Implementation Method 1
Many target nucleic acids are present in an organism in such low concentration, that a direct detection in a sample derived from that organism is not possible. Such targets need to be amplified before detection. Suitable amplification methods are for example LCR, cycling probe technology, Invader technology, Q-Beta replicase technology, NASBA, TMA, SDA and PCR
Implementation Method 2
The determination of nucleic acids has become an important tool in analytical chemistry, especially in health care. For example, infection diseases and genetic status can be easily determined on the basis of the presence or the amount of a nucleic acid indicative of said disease or status in samples received from the individual. For this reason methods were established using sequence specific hybridization of a nucleic acid, preferably an oligonucleotide, with a target nucleic acid
Data Source
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AI summary
The present invention is related to a method for detecting a target biomolecule in test sample by adding an internal control biomolecule to the test sample; to a negative control sample, to a positive control sample and to a reagent control sample or adding an internal control biomolecule to the test sample, to a negative control sample, to a positive control sample comprising the target biomolecule and providing a reagent control sample comprising the target biomolecule, determining in each sample a signal, and verifying the signal thereby detecting the target biomolecule. The invention is also related to a method for verifying the determination of a signal indicating the presence of a target biomolecule. The invention is further related to a method for detecting the presence or the absence of a member of a group of target nucleic acids in a sample and a method for verifying the determination of a signal indicating the presence of a member of a group of target nucleic acids. Uses and kits are considered as well.