One-Component Primer Probe Mixture for Coronavirus Diagnosis
Find Innovative SolutionsGenerate Solutions
Solution Overview
Problem
Current coronavirus diagnostic kits using real-time PCR face challenges with reliability due to the need to handle multiple containers for primer pairs and probes, leading to contamination and interference issues during virus diagnosis.
Innovation Solution
A one-component primer/probe mixture is developed, combining specific primer pairs and probes for amplifying coronavirus genes, including RdRp, N, E, and human RNase P genes, to enhance diagnostic reliability and convenience by reducing container handling and minimizing contamination risks.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If multiple primer pairs and probes are placed in separate containers to amplify specific viral RNA sites, then amplification reliability is improved, but ease of operation deteriorates due to difficulty in handling multiple containers simultaneously
Solution Approach 1:
The patent combines multiple primer pairs and probes that were previously stored in separate containers into a single integrated container. This merging allows all necessary reagents for amplifying different viral RNA sites to be handled simultaneously without the operational difficulty of managing multiple separate containers, while maintaining the reliability of specific amplification through carefully designed primer and probe sequences.
2Ease of operation
If multiple primer pairs and probes are randomly mixed to simplify handling, then ease of operation is improved, but analysis reliability deteriorates due to interference between primers
Solution Approach 1:
The patent applies local quality by designing each primer pair and probe with specific sequence characteristics optimized for their target viral RNA sites. Rather than randomly mixing primers, each component has tailored properties (specific base sequences, binding affinities, and target specificities) that ensure they function independently and specifically within the mixture, preventing interference while allowing convenient single-container handling.
Solution Approach 2:
The patent changes critical parameters of the primer and probe designs, including their base sequences, concentrations, and binding temperatures, to optimize their performance when mixed together. By carefully adjusting these parameters, the system achieves both ease of operation through single-container mixing and maintains analysis reliability by preventing primer-dimer formation and cross-interference.
3Measurement precision
If multiple containers are handled simultaneously to maintain separate primer pairs and probes, then diagnostic precision is improved, but contamination risk increases leading to degraded reliability
Solution Approach 1:
The patent merges multiple separate containers into a single container holding all primer pairs and probes. This eliminates the need to handle multiple containers simultaneously, thereby reducing the risk of cross-contamination between reagents. The single-container approach maintains diagnostic precision through carefully designed specific primers and probes while improving reliability by preventing contamination-related errors.
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 one-component coronavirus diagnostic kit improves analytic specificity, sensitivity, reproducibility, and repeatability, allowing for accurate and convenient detection of coronavirus with reduced interference, as demonstrated by experimental results showing high sensitivity and specificity.
Implementation Method 1
a coronavirus diagnostic kit using a real-time polymerase chain reaction (PCR) is used. The diagnostic kit using real-time PCR is based on a method of amplifying and confirming only a specific gene by extracting viral RNA from a collected sample and then performing reverse transcription of the extracted viral RNA.
Implementation Method 2
The diagnostic kit using real-time PCR is based on a method of amplifying and confirming only a specific gene by extracting viral RNA from a collected sample and then performing reverse transcription of the extracted viral RNA.
Data Source
AI summary
Provided are a method of manufacturing a coronavirus diagnostic kit, which includes preparing a one-component primer/probe mixture by mixing a primer set including a primer pair consisting of base sequences of SEQ ID NOs: 1 and 2, a primer pair consisting of base sequences of SEQ ID NOs: 3 and 4, a primer pair consisting of base sequences of SEQ ID NOs: 5 and 6, and a primer pair consisting of base sequences of SEQ ID NOs: 7 and 8; and a probe consisting of a base sequence of SEQ ID NO: 9, a probe consisting of a base sequence of SEQ ID NO: 10, a probe consisting of a base sequence of SEQ ID NO: 11, and a probe consisting of a base sequence of SEQ ID NO: 12, a coronavirus diagnostic kit manufactured using the same, and a method of diagnosing coronavirus using the same.