GeXP Multiplex PCR Primer Set for Avian Influenza Subtype Detection
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Solution Overview
Problem
Current methods for detecting avian influenza virus subtypes are complex, time-consuming, and lack the ability to simultaneously identify multiple subtypes, particularly the eight different human-infected subtypes H1, H2, H3, H5, H6, H7, H9, and H10, which are essential for rapid diagnosis and epidemiological investigation.
Innovation Solution
A GeXP multiplex PCR technique using a specific primer set and universal primer pair, optimized for the GenomeLab GeXP genetic analysis system, allows for the simultaneous detection of eight different human-infected subtypes of avian influenza virus in a single PCR reaction, providing high throughput, specificity, and sensitivity.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If virus isolation and culture method is used for diagnosis, then detection accuracy is improved, but detection time increases and operation complexity increases
Solution Approach 1:
The patent replaces the traditional mechanical virus isolation and culture method with a molecular biology-based GeXP multiplex PCR system. This substitution enables simultaneous detection of multiple avian influenza virus subtypes through genetic amplification and capillary electrophoresis, achieving both high accuracy and rapid results within 4 hours without the time-consuming culture process.
2Productivity
If common multiplex PCR is used for simultaneous amplification, then detection efficiency is improved, but detection accuracy decreases due to different amplification efficiency
Solution Approach 1:
The patent optimizes multiple parameters of the multiplex PCR system including primer concentrations, annealing temperatures, and Mg2+ concentrations to achieve balanced amplification efficiency across all target subtypes. The GeXP system uses fluorescently labeled primers and capillary electrophoresis with optimized separation conditions to ensure accurate differentiation and quantification of multiple amplification products simultaneously.
3Adaptability or versatility
If detection method is expanded to cover more subtypes, then adaptability is improved, but device complexity increases
Solution Approach 1:
The patent designs a universal GeXP multiplex PCR system that can simultaneously detect eight different human-infected avian influenza virus subtypes (H1N1, H2N2, H3N2, H5N1, H6N1, H7N9, H9N2, H10N8) using a single reaction体系. The system employs subtype-specific primers with universal fluorescent labels, allowing one detection platform to perform multiple subtype identification functions without requiring separate assays for each subtype.
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 method enables rapid detection and genotyping of eight avian influenza virus subtypes within four hours, enhancing epidemiological investigation, differential diagnosis, and public health safety by ensuring timely and accurate identification of human-infected subtypes.
Implementation Method 1
A GeXP multiplex PCR technique using a specific primer set and universal primer pair, optimized for the GenomeLab GeXP genetic analysis system, allows for the simultaneous detection of eight different human-infected subtypes of avian influenza virus in a single PCR reaction
Implementation Method 2
Multiplex reverse transcription polymerase chain reaction (mRT-PCR) in GeXP multiplex gene genetic expression analysis system can solve problem of different amplification efficiency present in the common multiplex PCR by combining universal primer (labeled with fluorescent marker upstream) and specific chimeric primer
Data Source
AI summary
Provided herein are a GeXP rapid detection primer set for simultaneously identifying gene HA of eight different human-infected subtypes of avian influenza virus, a kit, and use thereof. Disclosed are 9 pairs of specific primer and 1 pair of universal primer, for a GeXP rapid detection primer kit for simultaneously identifying gene HA of eight different human-infected subtypes of avian influenza virus. Eight different human-infected subtypes HA of avian influenza virus from the nine genes of M, H1, H2, H3, H5, H6, H7, H9 and H10 can be identified simultaneously with a sensitivity of 102 copies/μL.