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Updated: Jul 12, 2025
Influenza A Virus Studies in a Mouse Model of Infection
Published on: September 7, 2017
Detection of Minute virus of mice strains in different cell lines: Implications for adventitious agent testing
Maria R Farcet1, Jens Modrof1, Gerhard Antoine1
1Global Pathogen Safety, Takeda Manufacturing Austria AG, Vienna, Austria.
Abstract:
Minute virus of mice (MMV) has contaminated biotechnological processes in the past and specific MMV testing is therefore recommended, if the production cell line is known to be permissive for this virus. Testing is widely done using cell-culture-based adventitious virus assays, yet MMV strains may differ in their in vitro cell tropism. Here, we investigated the growth characteristics of different MMV strains on A9 and 324K cells and identified significant differences in susceptibility of these widely used indicator cell lines to infection by different strains of MMV, which has implications for MMV detectability during routine testing of biotechnology process harvests. An MMV-specific polymerase chain reaction was evaluated as a more encompassing method and was shown as suitable replacement for cell culture-based detection of the different MMV strains, with the additional benefit that detection is more rapid and can be extended to other rodent parvoviruses that might contaminate biotechnological processes. Although no MMV contamination event of human-derived cell lines has happened in the past, biotechnological processes that are based on these also need to consider MMV-specific testing, as, for example, HEK293, a human-derived cell line commonly used in biopharmaceutical manufacturing, was shown as susceptible to productive MMV infection in the current work.
Insights
Minute virus of mice (MMV) testing is crucial for biotechnological processes. New research shows MMV strains vary in cell susceptibility, highlighting the need for improved detection methods like PCR for broader rodent parvovirus coverage.
Area of Science:
- Virology
- Biotechnology
- Cell Biology
Background:
- Minute virus of mice (MMV) has previously contaminated biotechnological processes.
- Current MMV testing relies on cell-culture-based adventitious virus assays.
- MMV strains exhibit varying in vitro cell tropism, potentially impacting detection.
Purpose of the Study:
- To investigate the growth characteristics of different MMV strains on A9 and 324K cells.
- To assess the implications of MMV strain variability on detectability in biotechnology processes.
- To evaluate an MMV-specific polymerase chain reaction (PCR) as an alternative detection method.
Main Methods:
- Comparative analysis of MMV strain growth on A9 and 324K cell lines.
- Assessment of MMV susceptibility in commonly used indicator cell lines.
- Evaluation of an MMV-specific PCR assay for sensitivity and scope.
Main Results:
- Significant differences were identified in the susceptibility of A9 and 324K cells to various MMV strains.
- Human-derived cell lines, including HEK293, demonstrated susceptibility to productive MMV infection.
- The MMV-specific PCR assay proved effective for detecting diverse MMV strains and other rodent parvoviruses.
Conclusions:
- MMV strain variability necessitates careful consideration for cell-culture-based detection in biotechnology.
- MMV-specific PCR offers a more rapid, sensitive, and broader detection method for contaminants.
- Biotechnological processes using human-derived cell lines require MMV-specific testing due to demonstrated susceptibility.
