Influenza Detection via PB1 Nucleic Acid Amplification
Find Innovative SolutionsGenerate Solutions
Solution Overview
Problem
Current methods for detecting influenza viruses are inadequate for identifying both known and novel strains, particularly in animal hosts like bats, which can serve as reservoirs for zoonotic transmission to humans, and often fail to detect influenza-related viruses effectively.
Innovation Solution
The development of methods and compositions that amplify and detect influenza virus polymerase basic protein 1 (PB1) nucleic acid using specific primers capable of hybridizing to sequences at least 70% identical to known PB1 nucleic acid sequences, allowing for the detection of any type of influenza virus, including novel or unknown strains, in a sample.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If pathogen-specific assays are used to detect influenza viruses, then detection specificity for known strains is improved, but detection capability for novel and unknown influenza viruses deteriorates
Solution Approach 1:
The patent develops universal primers that can detect multiple types of influenza viruses (A, B, and C) and related viruses through a single assay. The primers are designed to target conserved regions of the PB1 nucleic acid that are present across different influenza virus types, enabling one assay to perform multiple detection functions rather than requiring separate pathogen-specific assays for each virus type.
Solution Approach 2:
The patent modifies the detection parameters by using degenerate primers with reduced stringency conditions, allowing the primers to bind to nucleic acid sequences with up to 30% divergence from known sequences. This parameter change enables detection of novel viruses that would not be detected by conventional high-stringency pathogen-specific assays, while still maintaining sufficient specificity through the conserved PB1 target region.
2Ease of operation
If conventional detection methods are used, then assay simplicity is maintained, but detection sensitivity for low viral loads deteriorates
Solution Approach 1:
The patent replaces conventional mechanical detection methods with nucleic acid amplification technology (PCR). Instead of directly detecting viral particles or antigens, the method amplifies viral nucleic acid to detectable levels, significantly improving detection sensitivity for low viral loads while maintaining operational simplicity through standardized PCR protocols that are widely available in clinical laboratories.
3Adaptability or versatility
If multiple pathogen-specific assays are implemented to detect all influenza virus types, then detection coverage is improved, but device complexity and cost increase
Solution Approach 1:
The patent merges the detection of multiple influenza virus types (A, B, and C) and related viruses into a single unified assay. By designing primers that target conserved regions common to all these viruses and using low-stringency hybridization conditions, the patent combines what would traditionally require multiple separate assays into one test, reducing device complexity and operational burden while maintaining comprehensive detection coverage.
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
These methods enable sensitive and specific detection of influenza viruses, including novel strains, with high sensitivity (100-500 copies) and specificity, facilitating early detection and monitoring of zoonotic viruses, thereby protecting public health.
Implementation Method 1
the at least one primer is capable of hybridizing to an influenza virus PB1 nucleic acid
Implementation Method 2
amplifying the PB1 nucleic acid to produce an amplified PB1 nucleic acid
Data Source
AI summary
Disclosed herein are methods of detecting influenza virus in a sample from a subject. In some embodiments, the disclosed methods include contacting a sample with at least one primer 10-40 nucleotides in length wherein the at least one primer is capable of hybridizing to an influenza virus polymerase basic protein 1 (PB1) nucleic acid at least 70% identical to the nucleic acid sequence set forth as any one of SEQ ID NOs: 1-3, amplifying the PB1 nucleic acid or a portion thereof to produce an amplified PB1 nucleic acid, and detecting the amplified PB1 nucleic acid, wherein presence of the amplified PB1 nucleic acid indicates presence of influenza virus in the sample from the subject. In some examples, the primers comprise or consist of the nucleic acid sequence set forth as one of SEQ ID NO: 4, SEQ ID NO: 5, SEQ ID NO: 9, or SEQ ID NO: 10.