HBV Detection Oligonucleotide Probes and Lyophilized Reagents
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Solution Overview
Problem
Current HBV detection methods face challenges due to significant genetic diversity, requiring improved reliability, reduced storage needs, and faster sample processing times, while also being prone to contamination and under-quantification of rare variants.
Innovation Solution
A set of oligonucleotide sequences, including forward and reverse primers, and dual-probe designs with detectable labels, is provided for amplifying and detecting HBV nucleic acid sequences, enhancing detection reliability and reducing storage requirements through lyophilized reagents and rapid sample-to-result analysis.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If a single probe is used for HBV DNA detection, then the assay is simpler and cheaper, but it results in under-quantification or lack of detection of rare HBV variants due to mutations within the probe region
Solution Approach 1:
The patent divides the detection system into multiple probes (at least two probes) that target different regions of the HBV genome. Each probe is designed to detect specific HBV variants, and their combined use ensures comprehensive detection across diverse genotypes and mutations, resolving the contradiction between detection reliability and assay complexity.
2Productivity
If liquid format PCR reagents are used, then the assay can be performed, but frozen storage is required and batch testing is needed which increases turnaround time
Solution Approach 1:
The patent changes the physical state of the PCR reagents from liquid to lyophilized (freeze-dried) form. This parameter change eliminates the need for frozen storage, allows individual sample processing without batch testing, and reduces turnaround time while maintaining reagent stability and functionality.
3Productivity
If pooled samples are tested, then throughput is increased, but DNA levels can drop below the limit of detection when the initial peak of virus resolves
Solution Approach 1:
The patent implements a dynamic detection approach using multiple probes with different target regions and amplification efficiencies. This dynamic system adapts to varying DNA concentrations in pooled samples, maintaining detection sensitivity across different viral load levels and preventing false negatives when virus levels fluctuate during the assay.
4Productivity
If conventional PCR methods are used, then standard protocols can be followed, but handling errors such as contamination occur and sample preparation takes several hours
Solution Approach 1:
The patent employs a self-contained lyophilized reagent system where all necessary PCR components are pre-packaged in a stable, contamination-free state. The lyophilized format eliminates the need for separate sample preparation steps and reduces handling operations, thereby minimizing contamination risks while enabling rapid processing.
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 oligonucleotide sequences enable reliable detection of HBV across multiple genotypes, reducing contamination risks and turnaround time, with the dual-probe design improving detection accuracy and minimizing storage needs, facilitating efficient HBV diagnosis.
Implementation Method 1
a first probe oligonucleotide sequence comprising SEQ ID NO: 3; and (d) a second probe oligonucleotide sequence comprising SEQ ID NO: 4
Implementation Method 2
each of the first and second probe oligonucleotide sequences comprises a detectable label
Data Source
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AI summary
The disclosure is directed to methods, kits, and compositions, for amplifying and detecting a human hepatitis B virus (HBV) in a sample, which comprises a variety of combinations of forward oligonucleotide primers, reverse oligonucleotide primers, and oligonucleotide probes.