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Mycoplasma removal: simple curative methods for viral supernatants
C Baronti1, B Pastorino, R Charrel
1Aix Marseille Univ, IRD French Institute of Research for Development, EHESP French School of Public Health, UMR_D 190 Emergence des Pathologies Virales, 13005 Marseille, France. Cecile.baronti@univ-amu.fr
This study presents two simple, effective methods for removing mycoplasma contamination from virus stocks. These protocols successfully decontaminate cell culture supernatants without affecting viral infectivity, ensuring high-quality virus collections.
Area of Science:
- Virology
- Microbiology
- Quality Management
Background:
- Maintaining high-quality virus collections is crucial for research, as exemplified by the European Virus Archive (EVA) FP7 infrastructure.
- Mycoplasma contamination in cell cultures poses a significant threat to virus stock integrity and experimental reproducibility.
- Existing mycoplasma elimination methods primarily focus on cell lines, with limited documentation for decontaminating valuable cell culture supernatants.
Purpose of the Study:
- To improve mycoplasma detection and eradication in cell culture supernatants stored at -80°C or freeze-dried.
- To establish standardized decontamination procedures for virus stocks to meet European Virus Archive quality standards.
- To evaluate the efficacy and impact on viral infectivity of two optimized mycoplasma removal methods.
Main Methods:
- Testing a diverse panel of mycoplasma-contaminated virus stocks, including enveloped and non-enveloped, RNA and DNA viruses.
- Applying two optimized, easy-to-perform decontamination protocols: one using Plasmocin™ and another using chloroform.
- Assessing the complete removal of mycoplasma from cell culture supernatants.
Main Results:
- Both Plasmocin™ and chloroform treatments successfully eradicated mycoplasma from all tested virus stocks.
- The decontamination procedures were effective for various virus types (enveloped/non-enveloped, RNA/DNA).
- Crucially, neither method compromised the infectivity of the treated viral samples.
Conclusions:
- Optimized Plasmocin™ and chloroform treatments provide effective and simple solutions for mycoplasma decontamination of virus stocks.
- These methods ensure the quality and integrity of valuable virus collections without loss of viral infectivity.
- The study provides essential, documented procedures for decontaminating cell culture supernatants, addressing a gap in current practices.
