GLuc-E2 Fusion Antigen for Rapid Porcine GETV Detection
Find Innovative SolutionsGenerate Solutions
Solution Overview
Problem
Current diagnostic methods for porcine Getah virus (GETV) are cumbersome, require specialized equipment, are susceptible to cross-contamination, and lack sensitivity, leading to misdiagnosis and inadequate prevention.
Innovation Solution
A luciferase immunoprecipitation system (LIPS) using a Gaussia luciferase (GLuc)-E2 fusion protein, prepared by recombinant expression in mammalian cells, for rapid and sensitive detection of GETV through luminescence intensity.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If RT-PCR method is used for porcine GETV detection, then detection capability is provided, but operational procedures become cumbersome and equipment requirements increase
Solution Approach 1:
The patent replaces the complex RT-PCR mechanical system with a luminescence-based detection system. The LIPS method uses luminescent bacteria or enzymes that emit light signals when interacting with GETV antigens, eliminating the need for complex thermal cycling, DNA amplification, and specialized PCR equipment, thereby simplifying operational procedures while maintaining detection capability
Solution Approach 2:
The patent changes the detection parameter from nucleic acid amplification (RT-PCR) to luminescence intensity measurement. By using luminescent markers that produce measurable light signals, the system achieves detection without requiring the complex mechanical and thermal processes of PCR, thus reducing device complexity while preserving measurement precision
2Measurement precision
If RT-PCR method is used for porcine GETV detection, then detection capability is provided, but cross-contamination risk increases
Solution Approach 1:
The patent replaces the RT-PCR system with a luminescence-based assay that does not involve DNA amplification or thermal cycling. The LIPS method uses direct antigen-antibody interactions followed by luminescent signal detection, eliminating the mechanical processes that create contamination risks in PCR laboratories, thereby reducing cross-contamination risk while maintaining detection capability
Solution Approach 2:
The patent employs disposable luminescent substrates and single-use reagent components that eliminate the need for complex, reusable PCR equipment. The luminescent signals are generated in vitro and do not require contamination-prone amplification machinery, reducing cross-contamination risk while preserving detection accuracy
3Measurement precision
If ELISA method is used for porcine GETV detection, then serological detection is achieved, but sensitivity and specificity depend on antigen quality
Solution Approach 1:
The patent introduces luminescent bacteria or enzymes as intermediary components that amplify and enhance the detection signal. Instead of relying solely on the quality of the GETV antigen itself, the system uses luminescent intermediaries that produce measurable signals when interacting with viral antigens, thereby improving sensitivity and reducing dependence on perfect antigen quality while maintaining specificity
Solution Approach 2:
The patent changes the detection parameter from direct antigen-antibody binding (ELISA) to luminescence signal generation. By using luminescent markers that emit light in response to viral antigens, the system achieves higher sensitivity and reduces the stringency requirements for antigen quality, while maintaining specificity through targeted luminescent signal detection
4Measurement precision
If ELISA method is used for porcine GETV detection, then serological detection is achieved, but operational complexity increases
Solution Approach 1:
The patent replaces the ELISA system with a luminescence-based detection method that simplifies operational steps. The LIPS method uses direct luminescent signal generation without requiring complex enzyme conjugations, substrate additions, or wash steps, thereby reducing operational complexity while maintaining or improving specificity through enhanced luminescent signal-to-noise ratios
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 LIPS method offers high sensitivity, specificity, and stability with a 100% coincidence rate with immunofluorescence detection, enabling rapid results within 3 hours and low operational complexity.
Implementation Method 1
A luciferase immunoprecipitation system (LIPS) can be utilized for porcine GETV infection detection based on luminescence intensity
Implementation Method 2
the fusion antigen is a Gaussia luciferase (GLuc)-E2 fusion protein
Data Source
AI summary
Provided are a fusion antigen of porcine Getah virus (GETV), a kit, a preparation method therefor and an application thereof. The fusion antigen of the GETV is primarily prepared by recombining a Gaussia luciferase (GLuc) gene with a codon- optimized GETV E2 antigen gene to construct an expression vector, and transfecting the expression vector containing GLuc-E2 into mammalian cell lines, resulting in the secretion of GLuc-E2 proteins into a cell supernatant for expression. Without the need for protein purification step, the cell supernatant may be directly collected for disease detection. The present disclosure demonstrates strong specificity and shows no cross- reactivity with African swine fever virus (ASFV), porcine reproductive and respiratory syndrome virus (PRRSV), porcine circovirus type 2 (PCV2), pseudorabies virus (PRV), or Japanese encephalitis virus (JEV).