Norovirus Detection Primer Set Targeting Intergenic Regions
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Solution Overview
Problem
Current methods lack primers capable of amplifying target sequences from the intergenic regions between ORF1 and ORF2, and ORF2 and ORF3 in norovirus, as well as specific probes for detecting these regions, which are crucial for sensitive and rapid norovirus detection.
Innovation Solution
Development of an oligonucleotide primer set and probe set that target the intergenic regions between ORF1 and ORF2, and ORF2 and ORF3 in norovirus, designed from highly conserved sequences found in 100 norovirus isolates, allowing for amplification and detection of norovirus using PCR and microarray hybridization.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Adaptability or versatility
If conventional primer sets are used for norovirus detection, then detection can be performed with existing methods, but they cannot amplify target sequences from intergenic regions between ORF1-ORF2 and ORF2-ORF3
Solution Approach 1:
The invention divides the norovirus detection task into multiple segments by designing separate primer sets for different intergenic regions (ORF1-ORF2 and ORF2-ORF3). This segmentation allows each primer set to be optimized for specific target sequences, enabling comprehensive coverage of norovirus genetic material that conventional single-purpose primers cannot achieve.
Solution Approach 2:
The primer sets are designed to be universal across different norovirus strains by targeting highly conserved intergenic regions. These primers can amplify target sequences from multiple norovirus genotypes simultaneously, providing multi-functional detection capability that works across diverse viral variants without requiring strain-specific primers.
2Measurement precision
If specific probes for intergenic regions are developed, then detection sensitivity increases, but the complexity of primer and probe design increases
Solution Approach 1:
The invention performs preliminary identification of highly conserved sequences in intergenic regions before designing primers and probes. By pre-characterizing the genetic conservation patterns across norovirus strains, the design process is simplified and detection sensitivity is maximized, as the targets are predetermined to be both sensitive and universally applicable.
Solution Approach 2:
The intergenic regions serve as intermediary target sequences that are more conserved and accessible than coding regions. These intermediary regions facilitate primer and probe binding while maintaining high detection sensitivity, acting as mediators between the variable viral genome and the detection system.
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 sensitive and specific detection of norovirus with high efficacy, capable of amplifying and hybridizing with target sequences, facilitating rapid and accurate identification of norovirus isolates.
Implementation Method 1
an oligonucleotide probe or probe set specifically hybridizing with at least one target sequence of the genomic RNA of norovirus
Implementation Method 2
an oligonucleotide primer set for amplifying at least one target sequence of genomic RNA of norovirus
Data Source
AI summary
Provided are an oligonucleotide primer set for amplifying at least one target sequence of the genomic RNA of norovirus, an oligonucleotide probe or probe set specifically hybridizing with at least one target sequence of the genomic RNA of norovirus, a microarray immobilized with the probe or probe set, and a method of detecting norovirus using the probe or probe set.