HPV Detection Primer Probe Sets for Genotype Differentiation
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Solution Overview
Problem
Current methods for detecting high-risk human papillomavirus (HPV) types are inadequate as they fail to simultaneously detect multiple genotypes with high specificity and sensitivity, often experiencing cross-reactivity and inefficiencies in clinical workflows.
Innovation Solution
Development of specific primers and probes sets, including forward and reverse primers and probes, designed to amplify and detect HPV types 16, 18, 31, 33, 35, 39, 45, 51, 52, 56, 58, 59, 66, and 68, and human beta globin sequences, utilizing sequences such as SEQ ID NOs: 1-22, which allow for robust amplification and differentiation in a single reaction without cross-reactivity.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Productivity
If multiple HPV genotypes are detected in a single reaction, then productivity is improved, but measurement precision deteriorates due to cross-reactivity between HPV types
Solution Approach 1:
The patent divides the detection system into multiple probe sets, each targeting specific HPV genotype groups. Forward primers amplify common regions across HPV types, while genotype-specific probes (e.g., probes for types 16, 18, 31, 33, 35, 39, 45, 51, 52, 56, 58, 59, 66, and 68) provide differentiated detection. This segmentation allows multiple genotypes to be detected in one reaction while maintaining precision through specific probe binding.
Solution Approach 2:
The patent applies local quality by using genotype-specific probes with unique binding sequences for different HPV types. Each probe set is designed with specific complementarity to its target HPV genotype's unique regions, ensuring specific detection. The probes are labeled with different detectable labels (e.g., fluorophores) to distinguish between different HPV types detected in the same reaction.
2Reliability
If HPV detection sensitivity is increased, then reliability is improved, but device complexity increases due to multiple primer and probe sets required
Solution Approach 1:
The patent creates a universal detection system where a single set of forward primers can detect multiple HPV genotypes simultaneously. The forward primers are designed to bind to conserved regions across different HPV types, while the genotype-specific probes provide the differentiation. This multi-functionality allows one primer set to serve multiple detection purposes, reducing overall system complexity while maintaining high sensitivity.
Solution Approach 2:
The patent uses genotype-specific probes as intermediaries between the universal forward primers and the HPV DNA targets. The probes act as mediators that provide specific binding to different HPV genotypes while being detected through a common signal detection system. This intermediary approach maintains sensitivity for each genotype while simplifying the overall assay architecture.
3Reliability
If human beta globin is used as internal control, then reliability is improved for assay validity, but measurement precision deteriorates due to potential interference with HPV detection
Solution Approach 1:
The patent segments the detection system into separate detection channels for HPV and human beta globin. HPV detection uses forward primers and genotype-specific probes, while beta globin detection uses separate primers and probes. This segmentation allows the internal control to validate assay performance without interfering with HPV genotype detection, as each target is detected through distinct molecular interactions.
Solution Approach 2:
The patent applies local quality by using distinct probe sequences and detection labels for HPV and beta globin. The beta globin probes are designed to bind specifically to human beta globin sequences with high specificity, while HPV probes target HPV-specific regions. This differentiation ensures that the internal control measurement does not cross-react or interfere with HPV detection precision.
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 primer and probe sets enable efficient detection of multiple HPV genotypes with high clinical sensitivity and specificity, providing a robust and efficient workflow for HPV detection and differentiation, including the use of human beta globin as an internal control for assay validity.
Implementation Method 1
The primer and probe sets enable efficient detection of multiple HPV genotypes with high clinical sensitivity and specificity, providing a robust and efficient workflow for HPV detection and differentiation
Implementation Method 2
probes, including forward and reverse primers and probes, designed to amplify and detect HPV types 16, 18, 31, 33, 35, 39, 45, 51, 52, 56, 58, 59, 66, and 68
Data Source
AI summary
The present invention relates to primers, probes, primer sets, primer and probe sets, methods and kits for detecting human papillomaviruses, human beta globin sequences and human papillomaviruses and human beta globin sequences in a test sample.