期刊
JOURNAL OF VIROLOGICAL METHODS
卷 299, 期 -, 页码 -出版社
ELSEVIER
DOI: 10.1016/j.jviromet.2021.114327
关键词
Mycoplasma contamination; Blood agar; Viral stock; Mycoplasma detection
资金
- Progetto di Ricerca di Ateneo 2020
- University of Pisa [PRA_2020_32, PRA_2020_37]
- bando PRIN 2017, Progetto di Ricerca di Interesse Nazionale [2017KM79NN]
- POR-CreO FESR Toscana 2020, Project Tuscavia [5143348]
- I-GENE, In-vivo Gene Editing by Nanotransducers, European call identifier H2020-FETOPEN-2018-2020 [862714]
Mollicutes, including Mycoplasma and Acholeplasma, are parasitic bacteria that are naturally resistant to many antibiotics and often found as contaminants in cultured cells. Various methods exist to detect Mycoplasma infection in cell lines, but they typically require hours of hands-on work to perform.
Mollicutes (Mycoplasma and Acholeplasma) are parasitic bacteria that adhere to cellular surfaces, naturally resistant to many antibiotics and extremely small. They are often found as contaminants in cultured cells, where they go unnoticed. They may be present in viral stocks because they are present in supernatants of cells where cultured viruses are released. The best way to keep laboratories free of Mycoplasma is to discard infected cultures, but, as judged by the very common finding of Mycoplasma-contaminated cultures in many laboratories, this is not done as often as it should be. A possible reason is that most procedures recommended take as long as performing a simple experiment and many laboratories delay testing to save money and time. Indeed, many methods exist to detect Mycoplasma infection of cell lines, but they take at least a couple of hours of hands-on work, if not more. Here we describe a procedure to screen viral stocks and tissue cultures for Mycoplasma presence. It relies on isolation of Mycoplasma on ordinary horse blood agar directly from exhausted tissue culture supernatants and does not require experienced personnel or expensive equipment. It only requires minutes of hands-on work, and, for this, it may be useful for weekly screening of cultures. It yields semiquantitative results in roughly 5 days, which is the time that usually passes between one subculture passage of cells in vitro to another. Because of its simplicity, it may be useful for detecting Mycoplasma in viral stocks and for frequent screening of cultures in research laboratories.
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