Attenuator Oligonucleotides for Nucleic Acid Detection
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Solution Overview
Problem
Existing nucleic acid detection methods struggle to accurately quantify target sequences of interest in samples where highly abundant target sequences (HATs) overwhelm the signal, making it difficult to detect less abundant sequences.
Innovation Solution
The use of attenuator oligonucleotides that can replace or compete with detector oligonucleotides for highly abundant target sequences, reducing their ligation and amplification products to facilitate the detection of other target sequences by hybridizing with them, thereby attenuating the signal of HATs.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If detector oligonucleotides are used to detect target sequences in a sample, then the presence of target sequences can be detected, but highly abundant target sequences (HATs) produce excessive signal that overwhelms the detection of less abundant sequences
Solution Approach 1:
An attenuator oligonucleotide is introduced as an intermediary substance that selectively binds to highly abundant target sequences (HATs) to reduce their signal. The attenuator acts as a mediator between the detector oligonucleotides and HATs, preventing detectors from binding to HATs and thereby reducing the overwhelming signal from abundant sequences while allowing detection of less abundant sequences to proceed normally
Solution Approach 2:
The signal parameter of highly abundant target sequences is changed by introducing attenuators that bind to HATs and prevent detector binding. This parameter change reduces the effective signal intensity of HATs from an overwhelming level to a manageable level that allows co-detection of less abundant sequences, while maintaining the detection capability for all target sequences
2Adaptability or versatility
If the sample contains both highly abundant and less abundant target sequences, then comprehensive detection coverage is achieved, but the dynamic range of the assay is exceeded making it difficult to detect less abundant sequences
Solution Approach 1:
The attenuator oligonucleotide serves as a selective intermediary that distinguishes between highly abundant and less abundant target sequences. By binding specifically to HATs, the attenuator creates a differential effect that preserves the detection signal for less abundant sequences while reducing the signal from abundant sequences, thereby improving signal discrimination within the assay
Solution Approach 2:
The attenuator creates a complementary binding relationship with HATs that mirrors the detector oligonucleotide binding but with opposite effect. While detectors amplify the signal from target sequences, the attenuator copies the binding interaction to suppress the signal from HATs, enabling the assay to handle the full dynamic range of target sequence abundances
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 allows for the detection of less abundant target sequences while maintaining a reduced signal from highly abundant ones, optimizing the dynamic range of the assay and enhancing the detection of target sequences of interest.
Implementation Method 1
The detectors are allowed to hybridize to nucleic acids in the sample. When downstream and upstream detectors both hybridize to the corresponding regions of the same target sequence
Data Source
AI summary
Methods for detecting nucleic acid sequences, where attenuator oligonucleotides are provided to reduce the number of detection products resulting from highly abundant sequences.