Multiplex RT-PCR Primer Set for Respiratory Virus Detection
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Solution Overview
Problem
Current methods for diagnosing viral respiratory diseases are limited by low sensitivity, risk of contamination, and long processing times, making early and accurate diagnosis challenging, especially for infants, elderly, and those with weakened cardiopulmonary function or immunity.
Innovation Solution
A primer set and composition for simultaneous diagnosis of viral respiratory diseases, comprising nucleotide sequences specific to 14 types of respiratory viruses, allowing for high sensitivity and specificity through real-time multiplex reverse transcription PCR, reducing contamination risks and processing time.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If conventional culture method is used for virus detection, then sensitivity is high, but testing time is too long (5-9 days) for early diagnosis
Solution Approach 1:
The patent extracts and amplifies only the specific viral gene sequences of interest using PCR, eliminating the need for lengthy cell culture processes. By targeting and amplifying specific viral genetic material directly from clinical samples, the method achieves both high sensitivity and rapid results within hours rather than days.
Solution Approach 2:
The patent replaces the mechanical cell culture system with a molecular biology-based PCR system. Instead of relying on cellular replication over several days, the invention uses enzymatic amplification of viral genes to achieve rapid detection, substituting a slow biological process with a faster biochemical reaction.
2Loss of time
If rapid antigen non-immunofluorescence test is used, then testing time is short, but sensitivity is low with high false negative results
Solution Approach 1:
The patent replaces antigen-antibody binding detection with nucleic acid amplification detection. By using PCR to amplify viral genetic material and detect it through fluorescence, the method achieves both rapid results and high sensitivity, overcoming the limitations of antigen-based rapid tests.
Solution Approach 2:
The patent changes the detection parameter from protein antigen levels to viral genetic material amplification. By measuring the accumulation of amplified DNA through fluorescence intensity, the method achieves superior sensitivity while maintaining rapid testing capability.
3Measurement precision
If molecular methods using electrophoresis are used, then sensitivity is good, but contamination risk increases due to two-step process and time consumption
Solution Approach 1:
The patent combines reverse transcription and PCR amplification into a single one-step reaction process. By integrating RNA-to-DNA conversion and subsequent amplification in one tube with continuous fluorescence monitoring, the method eliminates intermediate handling steps that could introduce contamination while maintaining high sensitivity.
Solution Approach 2:
The patent uses real-time fluorescence monitoring as an intermediary to detect viral presence during the amplification process itself, eliminating the need for post-PCR gel electrophoresis. This allows sensitivity detection without the contamination risks associated with opening amplified products.
4Measurement precision
If separate detection methods are used for different viruses, then specificity is high, but device complexity and processing time increase
Solution Approach 1:
The patent designs a universal PCR system with virus-specific primers and probes that can detect multiple different respiratory viruses simultaneously in a single reaction. By using fluorescently labeled probes with different emission wavelengths for different viruses, the method achieves multi-virus detection with a single integrated assay.
Solution Approach 2:
The patent adds the dimension of fluorescence wavelength differentiation to enable multiplexing. By assigning different fluorescent labels to different viral targets, the system can distinguish and detect multiple viruses simultaneously in the same reaction mixture, increasing throughput without proportionally increasing complexity.
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 prompt and accurate diagnosis of 14 types of respiratory viruses with high sensitivity and specificity, facilitating timely treatment and reducing the risk of unnecessary antibiotic use.
Implementation Method 1
real-time multiplex reverse transcription PCR
Implementation Method 2
real-time multiplex reverse transcription PCR
Implementation Method 3
nucleotide sequence selected from the group consisting of SEQ ID Nos. 1 to 14 and 17 to 33 for determining the infection of respiratory virus
Data Source
AI summary
The present invention relates to a kit for simultaneous diagnosis of viral respiratory diseases. To be more specific, the present invention is directed to a method for diagnosing viral respiratory diseases by detecting the genes specific to the respiratory disease-causing virus, a primer set for diagnosing the viral respiratory diseases used in the diagnosis method, a composition for simultaneous diagnosis of viral respiratory diseases, comprising the primer set, and a kit for simultaneous diagnosis of viral respiratory diseases, comprising the composition. When the primer set of the present invention for diagnosing the viral respiratory diseases is used, 14 different types of respiratory viruses can be simultaneously detected only with one reaction through real-time multiplex reverse transcription (RT)-PCR, and the onset of respiratory diseases caused by these viruses can be diagnosed. Thus, the primer set of the present invention can be widely used for prompt diagnosis and treatment of respiratory diseases.