HRV Detection via Specific Primers and Probes
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Solution Overview
Problem
Current methods for detecting human rhinovirus (HRV) in biological samples are inefficient due to high genetic variability and the presence of similar viral agents, leading to inaccurate and incomplete detection, especially in complex clinical or laboratory samples.
Innovation Solution
A process involving specific primers and probes targeting the 5′-noncoding region of HRV for PCR amplification, allowing for sensitive and discriminatory detection of HRV in biological samples, even in the presence of other viral agents, using techniques like real-time RT-PCR and mass spectrometry.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If conventional detection methods are used, then detection can be performed on various viral agents, but detection precision and reliability deteriorate due to high genetic variability and presence of similar viral agents
Solution Approach 1:
The invention divides the detection task into two distinct segments: first, a broad screening phase that captures multiple viral agents, and second, a specific identification phase that precisely distinguishes HRV from other viruses. This segmentation allows the system to maintain both versatility in detecting various viruses and precision in identifying HRV specifically.
Solution Approach 2:
The invention applies different detection qualities to different stages of the process. The initial screening uses general viral detection methods with broad applicability, while the subsequent confirmation stage employs highly specific primers and probes tailored specifically for HRV identification. This local differentiation of detection quality enables both versatility and precision.
2Adaptability or versatility
If broad viral detection methods are used, then various viral agents can be detected, but detection reliability worsens due to inability to distinguish HRV from similar agents
Solution Approach 1:
The detection process is segmented into a universal screening component that maintains versatility across viral agents, and a specific confirmation component that ensures reliability for HRV identification. The segmentation allows each component to optimize for its specific function without compromising the other.
Solution Approach 2:
The invention introduces specific HRV-targeting primers and probes as intermediary elements between the broad viral detection and final HRV identification. These intermediaries enable the system to transition from general viral detection to specific HRV confirmation, maintaining both versatility and reliability.
3Measurement precision
If non-specific PCR methods are used, then amplification can be performed on various viruses, but measurement precision deteriorates due to genetic variability
Solution Approach 1:
The invention applies specific quality characteristics to the PCR primers and probes, designing them to target highly conserved regions of the HRV genome. This localized optimization of primer specificity enhances detection sensitivity for HRV while maintaining manageable assay complexity through focused target selection.
4Reliability
If acid liability differentiation is used, then some HRV strains can be distinguished from HEVs, but detection precision worsens due to failure of some strains to grow in cell culture
Solution Approach 1:
The invention replaces the mechanical cell culture growth system with a direct molecular detection system using PCR. This substitution eliminates the productivity limitations of cell culture while maintaining or enhancing differentiation capability through sequence-specific amplification and detection methods.
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
This approach enables rapid, specific, and sensitive detection of HRV, distinguishing it from other viral pathogens, and provides quantitative data, improving diagnostic accuracy and reliability in clinical and laboratory settings.
Implementation Method 1
producing an amplification product by amplifying a human rhinovirus nucleotide sequence using a forward primer and a reverse primer and measuring the amplification product under conditions for a polymerase chain reaction
Implementation Method 2
The probe is illustratively of SEQ ID NO: 3... by hybridization of a probe complementary to a sequence from a human rhinovirus
Data Source
AI summary
A process for detecting human rhinovirus nucleic acid in a biological sample, includes producing an amplification product by amplifying an human bocavirus nucleotide sequence using a forward primer of SEQ ID NO: 1, and a reverse primer of SEQ ID NO: 2, and measuring said amplification product to detect human rhinovirus in said biological sample. Also provided are reagents and methods for detecting and distinguishing human rhinovirus from other viruses. A kit is provided for detecting and quantifying human rhinovirus in a biological sample.