Polynucleotide Primer Pairs for Simultaneous Pathogen Detection
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Solution Overview
Problem
Current diagnostic methods are inadequate for accurately detecting and distinguishing between hops latent viroid (HpLVd) and other pathogens like Beet Curly Top Virus (BCTV) in cannabis plants, leading to difficulties in assessing the relationship between pathogens and symptom presentation, which can result in reduced plant quality and productivity.
Innovation Solution
The development of methods and compositions involving nucleic acid analysis using specific polynucleotide primer pairs that hybridize and amplify subsequences of pathogens, allowing for the simultaneous detection of multiple pathogens, including HpLVd and BCTV, through techniques like RT-qPCR, enabling accurate identification and quantification of pathogens in plant samples.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If current diagnostic methods are used, then detection of pathogens can be performed, but accuracy in detecting and distinguishing between HpLVd and other pathogens like BCTV is inadequate
Solution Approach 1:
The diagnostic method is segmented into multiple independent PCR assays, each targeting specific pathogens (HpLVd, BCTV, AMV) with dedicated primer pairs. This segmentation allows each assay to be optimized for its specific target, improving detection accuracy while maintaining manageable complexity through modular design
Solution Approach 2:
Each primer pair is designed with local quality specific to its target pathogen's nucleic acid sequence. The primers exhibit high specificity to their intended target through careful sequence selection and optimization, enabling accurate distinction between different pathogens even when they co-infect the same plant
2Adaptability or versatility
If multiple pathogens are detected simultaneously, then comprehensive diagnosis is achieved, but the ability to distinguish between different pathogens and their genotypes becomes more difficult
Solution Approach 1:
The multiplex detection system is divided into separate, independent PCR reactions for each pathogen type. This segmentation prevents cross-reactivity and allows each assay to maintain high specificity for its target, enabling accurate differentiation even when detecting multiple pathogens simultaneously
Solution Approach 2:
The diagnostic approach dynamically adapts to different pathogen combinations by selecting and running only the necessary PCR assays for the sample being tested. This dynamic selection maintains precision by avoiding unnecessary assays while still providing comprehensive coverage when needed
3Reliability
If pathogen detection is performed to assess impact on plant quality, then disease management can be improved, but the relationship between pathogens and symptom presentation remains difficult to assess
Solution Approach 1:
The diagnostic method performs preliminary identification and quantification of pathogens before symptom analysis. By establishing which pathogens are present and at what levels prior to observing symptoms, the method creates a baseline that helps interpret symptom presentations and determine their causal relationships with detected pathogens
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
These methods provide accurate and reproducible diagnostics for detecting the presence, absence, and amount of pathogens, enabling effective assessment of pathogen impact on cannabis plants and improving disease management by distinguishing between different pathogens and their genotypes.
Implementation Method 1
specific polynucleotide primer pairs that hybridize and amplify subsequences of pathogens
Implementation Method 2
amplify subsequences of pathogens, allowing for the simultaneous detection of multiple pathogens, including HpLVd and BCTV, through techniques like RT-qPCR
Data Source
AI summary
The technology relates in part to methods and compositions for detecting one or more pathogens in plants. In some aspects, the technology relates to methods and compositions for detecting hops latent viroid in plants. In some aspects, the technology relates to methods and compositions for detecting hops latent viroid in cannabis plants. In some aspects, the technology relates to methods and compositions for classifying a hops latent viroid genotype. In certain aspects, the technology relates to methods and compositions for determining the presence, absence and/or amount of one or more pathogens in plants, either independently or simultaneously. In aspects, the pathogen is a virus. In some aspects, the virus is selected from among one or more of hops latent viroid, beet curly top virus and alfalfa mosaic virus.