Multiplex Immunoblot Assay for Early Veterinary Cancer Differentiation
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Solution Overview
Problem
Current cancer detection methods for pets, such as dogs, cats, and rabbits are inadequate for early detection, lack specificity, and are not economically accessible, particularly due to high sample volume requirements and inability to differentiate between cancer types.
Innovation Solution
A multiplex immunoblot and immunofluorescence assay is developed to detect onconeural/high-risk (ONHR) antibodies targeting intracellular neuronal antigens like amphiphysin, CV2, PNMa2/Ta, Ri, Yo, Hu, recoverin, SOX1, titin, Zic4, GAD65, and Tr (DNER) in serum or plasma samples, using nitrocellulose membrane strips and BIOCHIP slides, enabling early cancer detection and differentiation across various types.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If next-generation sequencing technology is used for early cancer detection in dogs, then sensitivity and specificity are improved, but sample volume requirement increases to 15-17 mL which is not suitable for medium and small dog breeds
Solution Approach 1:
The invention extracts and detects specific circulating tumor DNA fragments and onconeural antibodies from small blood samples (0.5-2 mL) rather than requiring large volumes for whole-genome sequencing. This extraction approach focuses on cancer-specific biomarkers present in circulation, enabling early detection in small breed dogs with minimal blood draw.
Solution Approach 2:
The invention changes the detection parameters from whole-genome sequencing to targeted detection of specific circulating tumor DNA fragments and onconeural antibody profiles. This parameter change allows highly sensitive cancer detection using significantly reduced sample volumes suitable for small and medium dog breeds.
2Measurement precision
If next-generation sequencing technology is used for early cancer detection, then detection accuracy is improved, but test cost increases making it unaffordable for pet owners with general and low income
Solution Approach 1:
The invention employs disposable, pre-prepared test cartridges containing capture probes and reagents for detecting circulating tumor DNA fragments and onconeural antibodies. These single-use cartridges eliminate the need for expensive next-generation sequencing equipment and infrastructure, making the test affordable for pet owners while maintaining high detection accuracy.
Solution Approach 2:
The invention replaces the complex mechanical and computational system of next-generation sequencing with a simplified immunological assay system using antibody-antigen binding and nucleic acid hybridization. This substitution dramatically reduces equipment costs and operational complexity while preserving cancer detection accuracy.
3Device complexity
If Nu.Q test based on nucleosome analysis is used for early cancer detection, then simplicity is improved, but ability to differentiate between cancer types and other conditions deteriorates
Solution Approach 1:
The invention segments the detection process into multiple independent assay channels: one for circulating tumor DNA fragment analysis and another for onconeural antibody profiling. Each segment targets specific cancer types and neurological conditions, allowing simultaneous differentiation of multiple cancer types and underlying causes while maintaining overall test simplicity.
Solution Approach 2:
The invention creates a multi-functional test platform that can detect various cancer types (lymphoma, osteosarcoma, mammary carcinoma, etc.), differentiate between malignant and benign conditions, and identify onconeural antibody-mediated neurological disorders all within a single integrated assay system.
4Measurement precision
If OncoK9 test is used for cancer detection, then sensitivity is improved, but ability to specify cancer type deteriorates except for lymphoma
Solution Approach 1:
The invention segments the detection panel into multiple cancer-type-specific biomarker assays, each targeting unique circulating tumor DNA mutations and onconeural antibody profiles associated with specific cancer types such as lymphoma, osteosarcoma, mammary carcinoma, and thyroid carcinoma, enabling precise cancer type specification.
Solution Approach 2:
The invention applies local quality by using different detection strategies optimized for specific cancer types: certain circulating tumor DNA mutations for solid tumors, onconeural antibody profiles for neuroendocrine tumors, and combined approaches for hematological cancers, thereby achieving both high sensitivity and specific cancer type identification.
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 assay allows for early cancer detection months before clinical symptoms, differentiates between 22 types of cancer and 16 neurological disorders, requires minimal sample volume, and is cost-effective, making it accessible to a broader range of pet owners.
Implementation Method 1
binding the composite specimen to at least one antigen selected from a group of antigens consisting of amphiphysin, CV2, PNMa2/Ta, Ri, Yo, Hu, recoverin, SOX1, titin, zic4, GAD65, and Tr (DNER), wherein the at least one antigen of the group of antigens is immobilized on a test strip
Implementation Method 2
optically detecting a presence of at least one antigen-antibody binding complex using a predetermined threshold
Implementation Method 3
A multiplex immunoblot and immunofluorescence assay is developed to detect onconeural/high-risk (ONHR) antibodies
Data Source
AI summary
The invention provides a qualitative immunoblot-based in vitro method for the detection of onconeural antibodies class IgG to twelve different antigens (amphiphysin, CV2, PNMa2/Ta, Ri, Yo, Hu, recoverin, SOX1, titin, zic4, GAD65 and Tr (DNER)) in serum or plasma samples of mammalian animals such as dogs, cats, ferrets, and rabbits for early diagnosis of twenty two cancer and cancer-associated neurological diseases. Detection of these antibodies in the blood of the animals is confirmed via an indirect immunofluorescent assay. Examples, including enzyme anti-dog, anti-cat, anti-ferret, and anti-rabbit conjugates, and serum or plasma quantities, are provided.