2016
DOI: 10.1002/cpmc.2
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Mimiviruses: Replication, Purification, and Quantification

Abstract: The aim of this protocol is to describe the replication, purification, and titration of mimiviruses. These viruses belong to the Mimiviridae family, the first member of which was isolated in 1992 from a cooling tower water sample collected during an outbreak of pneumonia in a hospital in Bradford, England. In recent years, several new mimiviruses have been isolated from different environmental conditions. These giant viruses are easily replicated in amoeba of the Acanthamoeba genus, its natural host. Mimivirus… Show more

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Cited by 10 publications
(11 citation statements)
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“…After the appearance of a cytopathic effect and evidence of cell lysis, the cells and supernatants were collected and the viruses were purified through ultracentrifugation with a 22% sucrose cushion at 36,000 × g for 30 min. After viral purification, the virus titers were determined using the endpoint method 17,18 .…”
Section: Methodsmentioning
confidence: 99%
“…After the appearance of a cytopathic effect and evidence of cell lysis, the cells and supernatants were collected and the viruses were purified through ultracentrifugation with a 22% sucrose cushion at 36,000 × g for 30 min. After viral purification, the virus titers were determined using the endpoint method 17,18 .…”
Section: Methodsmentioning
confidence: 99%
“…Then, the supernatant was collected, added over a 40% sucrose (Merck, Germany) cushion and centrifuged at 36,000 x g for 1 h. The pellet was re-suspended in PBS and stored at − 20 °C. Three aliquots of the virus stock were titrated to the 50% end-point and calculated by the Reed-Muench method [7, 8].…”
Section: Methodsmentioning
confidence: 99%
“…The same multiplicity of infection (MOI = 1) was used for each virus. The MOI was determined using the TCID50 method, as previously described 42 . Uninfected cells served as negative controls.…”
Section: Methodsmentioning
confidence: 99%