Antibody-Based Detection of Citrus Pathogen Secreted Proteins
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Solution Overview
Problem
Current methods for detecting citrus stubborn disease (CSD) and Huanglongbing (HLB) in citrus plants are hindered by the low titer and uneven distribution of pathogens, making nucleic acid-based assays costly, time-consuming, and inefficient for large-scale sample processing, and there are no curative methods once trees are infected.
Innovation Solution
A method involving the detection of secreted proteins from Spiroplasma citri for CSD and Candidatus Liberibacter asiaticus for HLB using antibodies, which can identify the presence of these proteins in untreated sap samples without the need for protein separation or purification, allowing for rapid and economical processing of multiple samples.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If nucleic acid-based assays are used to detect pathogens, then detection accuracy can be achieved, but the cost and time consumption increase significantly when processing large numbers of samples
Solution Approach 1:
The invention extracts and detects pathogen-specific proteins directly from plant tissue samples using antibody-based assays. By targeting secreted proteins that are readily available in plant tissues rather than searching for rare pathogen cells or nucleic acids, the method achieves high detection accuracy while dramatically reducing processing time and cost for large sample sets.
Solution Approach 2:
The invention uses antibodies as intermediaries to detect pathogen presence. Instead of directly detecting rare pathogen cells or nucleic acids, the method employs antibody-protein interactions to amplify and facilitate detection of pathogen presence through readily available secreted proteins, thereby improving both accuracy and efficiency.
2Reliability
If nucleic acid-based assays are used for pathogen detection, then positive diagnosis can be obtained, but the complexity and cost of sample preparation increase
Solution Approach 1:
The invention extracts pathogen-specific secreted proteins directly from plant tissue for detection. This approach eliminates the need for complex nucleic acid extraction and purification steps, simplifying sample preparation while maintaining reliable positive diagnosis through antibody-based detection of pathogen proteins.
Solution Approach 2:
The invention leverages the natural secretion of pathogen proteins into plant tissues, allowing the pathogen to essentially provide its own detection target. This self-service approach eliminates the need for complex sample preparation to extract or concentrate pathogen material, as the secreted proteins are already readily available in the plant tissue for direct antibody detection.
3Measurement precision
If pathogen cells are targeted for detection, then specific identification can be achieved, but the detection fails when pathogen titer is low or distribution is uneven
Solution Approach 1:
The invention extracts and detects pathogen-specific secreted proteins that are distributed throughout plant tissues rather than relying on rare pathogen cells. This approach ensures consistent detection across samples with low or unevenly distributed pathogen titers, as the secreted proteins are more uniformly distributed and readily available for antibody binding.
Solution Approach 2:
The invention uses secreted proteins as intermediary targets that bridge the gap between rare pathogen cells and detectable signals. These proteins serve as abundant, evenly distributed markers that maintain detection reliability while preserving pathogen-specific identification through antibody specificity.
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 approach enables robust and consistent detection of CSD and HLB, overcoming the limitations of pathogen distribution and providing a means for early disease management by identifying infected plants before symptoms develop.
Implementation Method 1
detecting the presence or absence of a secreted protein from Spiroplasma citri in a sample from the plant
Data Source
AI summary
Secreted proteins as detection markers for insect vector and graft transmitted citrus disease are described. Method and kits for detecting the secreted proteins are provided.