Ma-104 Cell Line for African Swine Fever Virus Detection
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Solution Overview
Problem
Current methods for detecting African Swine Fever Virus (ASFV) in field samples rely on primary swine macrophages, which are time-consuming, labor-intensive, and not readily available, and real-time PCR cannot distinguish between infectious and non-infectious virus, posing challenges in outbreak management.
Innovation Solution
Utilization of a stable cell line derived from African green monkey cells (Ma-104) for ASFV isolation and detection, employing red blood cell rosetting and nucleic-acid-based methods to identify replicating virus, allowing for efficient detection comparable to primary swine macrophages.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If primary swine macrophages are used for ASFV detection, then detection sensitivity is maintained, but time consumption and labor intensity increase significantly
Solution Approach 1:
The patent applies preliminary action by pre-adapting ASFV to the Ma-104 cell line through multiple passages, creating a stable viral strain that can efficiently infect and replicate in this cell type. This pre-adaptation enables rapid detection without requiring time-consuming primary macrophage isolation, thus reducing detection time while maintaining sensitivity
Solution Approach 2:
The patent uses Ma-104 cells as a substitute copy of primary swine macrophages for virus detection purposes. The adapted virus-Ma-104 system replicates the detection functionality of primary macrophages but with the advantage of being a stable, readily available cell line that eliminates the need for fresh primary cell isolation each time
2Reliability
If primary swine macrophages are used for ASFV detection, then virus isolation capability is maintained, but availability and ease of operation deteriorate
Solution Approach 1:
The patent replaces the difficult-to-obtain primary swine macrophages with Ma-104 cells that have been adapted to support ASFV replication. This substitution maintains virus isolation capability while dramatically improving availability, as Ma-104 cells can be stored and used repeatedly without requiring fresh isolation from animal sources
Solution Approach 2:
The patent changes the cellular parameter from primary macrophages (temporary, hard to obtain) to a stable cell line (permanent, easily available). This parameter change transforms the detection system from one requiring frequent re-preparation to a stable, reproducible system that can be maintained in the laboratory
3Speed
If real-time PCR is used for ASFV detection, then detection speed is improved, but ability to distinguish infectious virus deteriorates
Solution Approach 1:
The patent merges the speed advantage of molecular methods with the functional assessment capability of cell culture. By using Ma-104 cells that can be rapidly infected and show visible cytopathic effects or hemadsorption, the system combines quick detection with the ability to confirm infectious virus, eliminating the limitation of PCR alone
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 Ma-104 cell line enables rapid and sensitive detection of infectious ASFV, comparable to primary swine macrophages, facilitating effective epidemiological management and reducing the reliance on scarce primary cell sources.
Implementation Method 1
contacting the sample with a culture of Ma-104 cells under conditions that allow for infection of the cells by ASFV; incubating the Ma-104 cell and sample mixture under conditions that allow for ASFV detection; analyzing the mixture from step b for the presence or absence of replicating ASFV
Implementation Method 2
the analyzing step of the method is performed by adding red blood cells to the sample/Ma-104 mixture, and examining the Ma-104 cells for the presence or absence of hemadsorbed red blood cells on the Ma-104 cells
Data Source
AI summary
Methods are provided herein utilizing a stable cell line capable of efficient infection by African swine fever virus (ASFV) and also provides for the detection of the presence of virus in samples applied to the cells. Detection of the virus by means such as red blood cell rosetting is a surprising result given that the cell line is derived from African green monkeys. This cell line provides a marked improvement over the currently available testing strategies.