Multiplex Virus Detection Using Solid-Phase PCR Optimization
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Solution Overview
Problem
Current virus detection methods in cell therapy are limited to single or few viruses from the same species, face challenges like nonspecific binding and variability in amplification efficiency, and require costly trial-and-error testing for reaction conditions.
Innovation Solution
A method involving coating nucleic acid primers on a solid-phase carrier, setting optimized amplification reaction conditions, and conducting a polymerase chain reaction to detect multiple viruses from different species simultaneously, using parameters like temperature, primer length, and cycle number, while identifying and optimizing conditions for highest sensitivity.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Productivity
If multiplex PCR technology is applied to detect multiple viruses simultaneously, then detection efficiency is improved, but nonspecific binding between primers and variability in amplification efficiency occur
Solution Approach 1:
The patent optimizes critical PCR parameters including setting primer concentration between 0.1-1.0 μM (preferably 0.5 μM), reaction temperature between 50-70°C (preferably 65°C), and cycle number between 20-40 (preferably 30 cycles). These parameter adjustments resolve the contradiction by establishing conditions that enable simultaneous detection of multiple viruses while maintaining specific binding and consistent amplification efficiency.
2Productivity
If current multiplex PCR methods are used, then several viruses can be detected, but suboptimal reaction conditions lead to false positives and negatives
Solution Approach 1:
The patent performs preliminary optimization of reaction conditions before actual virus detection. This includes pre-determining optimal primer concentrations, reaction temperatures, and cycle numbers through systematic experimentation. By establishing these optimal conditions in advance, the method achieves both high productivity in detecting multiple viruses and high precision with reduced false positives and negatives.
3Measurement precision
If traditional virus detection methods are applied, then single or limited viruses can be detected, but costly and time-consuming trial-and-error testing is required for reaction conditions
Solution Approach 1:
The patent develops a universal optimized PCR protocol that can simultaneously detect multiple viruses from different species (human, bovine, porcine, avian) using a standardized set of reaction conditions. This multi-functional approach eliminates the need for separate trial-and-error optimization for each virus, reducing time loss while maintaining high detection specificity through the universally optimized parameters.
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
Enables efficient, accurate, and cost-effective detection of multiple viruses from different species with reduced false positives/negatives, replacing trial-and-error testing and improving diagnostic efficiency.
Implementation Method 1
coating a nucleic acid primer sequence corresponding to each of the viruses on a solid-phase carrier
Implementation Method 2
conducting an amplification reaction on the sample input into the solid-phase carrier according to the amplification reaction conditions
Data Source
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AI summary
A method for virus detection is applied to detect a plurality of viruses at one time and the method comprises the following steps of: coating a nucleic acid primer sequence corresponding to each of the viruses on a solid-phase carrier, wherein the viruses originate from different species; inputting a sample into the solid-phase carrier; setting a plurality of amplification reaction conditions, wherein the amplification reaction conditions comprise an amplification reaction temperature, an amplification primer length, an amplification primer concentration and a number of amplification reaction cycles; and conducting an amplification reaction on the sample input into the solid-phase carrier according to the amplification reaction conditions, so as to detect the plurality of viruses originating from different species at one time under the amplification reaction.