Ebola Virus Detection Primers and Probes for Specific RNA Discrimination
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Solution Overview
Problem
Current diagnostic techniques for Ebola virus detection are inadequate due to their inability to provide rapid, accurate, and specific results without specialized laboratories or equipment, leading to challenges in early diagnosis and effective public health responses during outbreaks.
Innovation Solution
Development of specific primers and probes capable of hybridizing with Ebola virus RNA, allowing for sensitive and specific detection and discrimination between Ebola virus and other viral hemorrhagic fever viruses, using PCR-based methods that can be deployed in non-specialized settings.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If prior RT-PCR assays are used for Ebola virus detection, then sensitivity is improved, but false positive results occur due to conserved genomic sequences
Solution Approach 1:
The patent applies local quality by designing primers and probes that target specific variable regions within the Ebola virus genome rather than conserved regions. The forward primer targets the NP gene region while the reverse primer targets the L gene region, and the probe targets a specific intermediate sequence. This localized targeting of specific genomic regions with unique sequences enables discrimination between Ebola virus and other filoviruses, resolving the contradiction between maintaining detection sensitivity and achieving viral specificity.
2Measurement precision
If specialized laboratories and equipment are used for Ebola diagnosis, then detection accuracy is improved, but device complexity and operational difficulty increase
Solution Approach 1:
The patent replaces complex mechanical and biological diagnostic systems with a simplified molecular biology approach. Instead of requiring viral culture in BSL-4 laboratories or transmission electron microscopy with specialized equipment, the invention uses RT-PCR with specifically designed primers and probes that can be performed with standard molecular biology equipment. This substitution of the diagnostic mechanism maintains high detection accuracy while dramatically reducing device complexity and laboratory specialization requirements.
3Ease of operation
If antibody ELISAs are used for Ebola detection, then operational simplicity is improved, but reliability decreases due to unreliable immune response
Solution Approach 1:
The patent applies preliminary action by detecting viral genetic material (RNA) directly from the patient sample through RT-PCR amplification, rather than waiting for the patient's immune system to produce detectable antibody responses. This early detection of viral nucleic acids occurs during the acute phase of infection before the immune response is fully developed, thereby maintaining operational simplicity while significantly improving detection reliability in symptomatic patients.
4Productivity
If prior RT-PCR assays are used, then rapid field deployment is achieved, but false positives occur due to inability to distinguish Ebola from other VHF viruses
Solution Approach 1:
The patent applies segmentation by dividing the diagnostic task into two distinct components: (1) a screening PCR assay that rapidly detects the presence of any filovirus through conserved region amplification, and (2) a discrimination step using subtype-specific primers and probes that identify the particular Ebola virus species. This segmented approach maintains the rapid detection capability of PCR while adding specificity through targeted sequence discrimination, enabling both speed and accuracy in field deployment.
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 accurate detection of Ebola virus, including subtypes, reducing false positives and autoinfection risks, and facilitating timely treatment and public health measures.
Implementation Method 1
hybridizing an Ebola virus RNA to an Ebola specific probe
Implementation Method 2
amplifying the Ebola nucleic acids with at least one primer specific for an Ebola nucleic acid
Data Source
AI summary
This invention relates to primers and probes for detecting Ebola virus and one or more subtypes of Ebola virus as well as kits including the probes and primers and methods of using the probes and primers.