Adenovirus Nucleic Acid Detection via Hybridization Probes
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Solution Overview
Problem
Current methods for detecting Adenovirus are limited in their ability to specifically and sensitively detect multiple serotypes due to the low homology of Adenovirus DNA sequences, particularly in immunosuppressed patients, where traditional serological tests and cell culture methods are slow and unreliable.
Innovation Solution
A method using in vitro nucleic acid amplification techniques with specific amplification oligomers and detection probes that target regions within the Adenovirus genome, allowing for the sensitive and specific detection of multiple serotypes by generating amplicons and using stringent hybridization conditions.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If traditional serological tests and cell culture methods are used for Adenovirus detection, then the methods are simple and widely applicable, but the detection speed is slow and reliability is reduced in immunosuppressed patients
Solution Approach 1:
The patent replaces traditional mechanical/cellular detection methods (serological tests and cell culture) with molecular nucleic acid amplification techniques. This substitution enables rapid detection of Adenovirus DNA through PCR-based assays, achieving both high reliability and reduced detection time, particularly for immunosuppressed patients who cannot mount adequate immune responses.
Solution Approach 2:
The patent changes the detection parameter from detecting viral proteins or cellular responses to directly detecting viral nucleic acids. This parameter change allows for earlier detection before immune responses develop and provides more reliable results in immunosuppressed patients, while maintaining simplicity through standardized molecular biology protocols.
2Productivity
If PCR-based assays are used to detect Adenovirus, then detection speed and sensitivity are improved, but the ability to detect multiple serotypes is limited due to low DNA sequence homology
Solution Approach 1:
The patent designs universal primer pairs that can detect multiple Adenovirus serotypes through a single PCR assay. The primers are engineered to bind to conserved regions across different serotypes, enabling one assay to serve multiple detection functions and expanding serotype coverage without compromising detection speed.
Solution Approach 2:
The patent employs dynamic primer design where the primer sequences are optimized to accommodate sequence variations among different serotypes. This dynamic approach allows the PCR assay to adapt to different viral variants while maintaining rapid detection, balancing versatility with speed through careful primer sequence selection and annealing condition optimization.
3Adaptability or versatility
If low stringency conditions are used in PCR assays to detect diverse adenoviruses, then detection coverage is improved, but specificity is reduced leading to false positives
Solution Approach 1:
The patent optimizes the stringency parameters of the PCR assay, including annealing temperature, primer concentration, and extension conditions. By carefully tuning these parameters, the assay achieves high specificity to distinguish Adenovirus from other viruses while maintaining broad coverage across multiple serotypes, preventing false positives through precise parameter control.
Solution Approach 2:
The patent uses probe sequences that serve as copies or complements of specific viral regions. These probes hybridize to the amplified DNA with high specificity, allowing the assay to confirm Adenovirus detection while maintaining broad serotype coverage. The probes act as verification copies that ensure accurate identification without cross-reacting with non-target viruses.
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 rapid and specific detection of Adenovirus nucleic acid across various serotypes, improving diagnostic accuracy and speed, particularly in high-risk individuals, while minimizing exposure risks for laboratory personnel.
Implementation Method 1
contacting a sample suspected of containing at least an Adenovirus nucleic acid with at least two amplification oligomers for generating an amplicon, wherein each of said at least two amplification oligomers is from 10 to about 50 nucleotides in length and wherein the amplification oligomers are respectively configured to specifically hybridize to regions within a target sequence of Adenovirus
Data Source
AI summary
There is disclosed a method for detecting an Adenovirus nucleic acid comprising the steps of: (a) contacting a test sample comprising nucleic acid with at least one hybridization assay probe having a nucleotide sequence consisting of a sequence selected from the group consisting of SEQ ID NOs: 10, 19, 21, 23, 24, 29, 36, and 37 or a combination of two or more thereof; and (b) determining if a probe:target duplex has formed under stringent hybridization conditions in the test sample, wherein the presence of the probe:target duplex is an indication of the presence of Adenovirus nucleic acid in the test sample.