Molecular Combing Detection of Infectious HSV DNA
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Solution Overview
Problem
Current methods for detecting infectious viral DNA in infected cells or tissues are inefficient, particularly in biological fluids, and cannot reliably identify complete infectious viral genomes or monitor antiviral treatment efficacy.
Innovation Solution
Molecular Combing and DNA stretching techniques combined with specially designed probes allow for the detection and quantification of infectious viral DNA in biological samples, enabling the identification of viral genomes and monitoring of antiviral treatment effects by immobilizing and hybridizing nucleic acids with sequence-specific probes.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If conventional detection methods are used for infectious viral DNA, then detection can be performed, but detection efficiency and accuracy are insufficient particularly in biological fluids
Solution Approach 1:
The detection method segments the viral DNA detection process into distinct phases: extraction of viral particles from biological fluids, isolation of viral genomes, and hybridization with sequence-specific probes. This segmentation allows optimization of each step for maximum sensitivity and specificity, particularly improving detection in complex biological fluid matrices where conventional methods fail
Solution Approach 2:
The patent employs sequence-specific hybridization probes as intermediaries between the target viral DNA and the detection system. These probes act as mediators that selectively bind to complementary viral genome sequences, enabling highly specific detection of infectious viral DNA while distinguishing it from non-infectious forms or host DNA in biological samples
2Loss of information
If conventional methods are used, then detection is possible, but the ability to identify complete infectious viral genomes is limited
Solution Approach 1:
The hybridization probe system is designed with multi-functionality to simultaneously detect the presence of viral DNA, determine genome completeness, and identify specific viral strains. The probes can hybridize to various regions of the viral genome, providing comprehensive information about the infectious viral genome in a single detection assay
Solution Approach 2:
The patent utilizes colorimetric or fluorescent labels on hybridization probes that produce detectable color or fluorescence changes upon binding to target viral DNA sequences. This visual signal change enables precise identification of complete infectious viral genomes and distinguishes them from incomplete or non-infectious forms through the presence or absence of specific hybridization patterns
3Reliability
If conventional detection approaches are used, then viral presence can be detected, but monitoring of antiviral treatment efficacy is not reliable
Solution Approach 1:
The detection method performs preliminary characterization of viral DNA integrity and completeness before treatment evaluation. By establishing baseline genome completeness and infectious viral load using the hybridization probe system, clinicians can accurately monitor treatment efficacy over time and distinguish between reduction in viral load versus improvement in genome integrity
Solution Approach 2:
The patent implements a feedback mechanism where detection results provide quantitative information about viral genome completeness and infectious particle concentration. This feedback enables real-time assessment of antiviral treatment response, allowing clinicians to adjust therapy based on measured changes in infectious viral DNA levels and genome integrity markers
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 method provides a rapid, accurate, and sensitive detection of infectious viral polynucleotides, overcoming limitations of existing techniques by enabling simultaneous detection and monitoring of viral infections, including the identification of complete viral genomes and antiviral treatment efficacy.
Implementation Method 1
The probe is hybridized to a complementary polynucleotide sequence
Data Source
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Figure 2A~2B
Figure 3(1)A~3(1)B
AI summary
A method for detecting in vitro the presence of a genome of a DNA virus or a viral derived DNA in an infected eukaryotic cell, tissue or biological fluid using Molecular Combing or other nucleic acid stretching methods together with probes, especially nucleic acid probes, having a special design. A method for monitoring in vitro the effects of anti-viral treatment by following the presence of genomic viral or viral derived DNA polynucleotides in a virus-infected cell, tissue or biological fluid. Detection of an infectious form of a virus using Molecular Combing and DNA hybridization. A kit comprising probes used to carry out these methods and a composition comprising the probes.