EBV Nucleic Acid Detection Using Segmented Oligonucleotide Probes
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Solution Overview
Problem
There is a need for a simplified and accurate approach to detect Epstein-Barr virus (EBV) infection, as existing methods struggle to specifically and sensitively detect EBV without cross-reacting with other herpesvirus or human nucleic acids.
Innovation Solution
A reaction mixture and method are developed that include a detection probe oligomer and a pair of amplification oligomers specific to EBV nucleic acids. The detection probe is up to 30 nucleotides in length and includes the base sequence of SEQ ID NO:22 or its complement, while the amplification oligomers are 18-25 nucleotides in length and include sequences from SEQ ID NO:3 and SEQ ID NO:4. This allows for the specific amplification and detection of EBV nucleic acids in a sample.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If conventional detection methods are used to detect EBV, then detection can be performed, but cross-reactivity with other herpesvirus and human nucleic acids occurs, reducing specificity
Solution Approach 1:
The detection system is segmented into multiple specific oligonucleotide probes (detection probe and amplification oligomers) that each target specific EBV nucleic acid sequences. This segmentation allows the system to distinguish EBV from other herpesviruses and human nucleic acids, achieving high specificity while avoiding cross-reactivity issues.
2Measurement precision
If detection sensitivity is increased to detect low levels of EBV, then more copies can be detected, but the risk of detecting non-EBV nucleic acids increases
Solution Approach 1:
The system uses specific oligonucleotide probes as intermediaries that bind selectively to EBV nucleic acid sequences. The detection probe (SEQ ID NO:22) and amplification oligomers (SEQ ID NO:3 and SEQ ID NO:4) act as mediators that enable sensitive detection of low-copy EBV while maintaining reliability through their sequence-specific binding, preventing false detection of non-EBV nucleic acids.
3Measurement precision
If complex detection protocols are used to improve accuracy, then detection precision improves, but the complexity of the procedure increases
Solution Approach 1:
The detection and amplification functions are merged into a single reaction mixture containing the detection probe and amplification oligomers. This combination allows the system to achieve high detection accuracy through specific sequence recognition while simplifying the procedure by eliminating separate detection and amplification steps.
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 described method enables sensitive and specific detection of EBV, distinguishing it from other herpesviruses and human nucleic acids, with the ability to detect as few as 50 copies of EBV per milliliter in a sample within a reasonable time frame.
Implementation Method 1
a detection probe oligomer for detecting EBV nucleic acids
Implementation Method 2
performing an in vitro nucleic acid amplification reaction using the oligomer combination, where any EBV target nucleic acid, if present in the sample, is a template for generating an amplification product
Data Source
AI summary
Disclosed are compositions, methods, and kits that can be used to Epstein-Barr virus (EBV) in a sample undergoing testing. Nucleic acids of EBV can be isolated, amplified and detected with specificity, and without interference from non-EBV organisms. In some embodiments, nucleic acids used for amplification are isolated from human blood, or blood products. Nucleic acid isolation, amplification and detection steps can all be carried out using an automated instrument.