Abstract
Fruit bats and insectivorous bats are believed to provide a natural reservoir for a wide variety of infectious diseases. Several lines of evidence, including the successful isolation of infectious viruses, indicate that Marburg virus and Ravn virus have found a major reservoir in colonies of the Egyptian rousette (Rousettus aegyptiacus). To facilitate molecular studies on virus-reservoir host interactions and isolation of viruses from environmental samples, we established cell lines from primary cells of this animal. The cell lines were given to several laboratories until we realized that a contamination with Vero cells in one of the cultures had occurred. Here we describe a general diagnostic procedure for identification of cross-species contamination with the focus on Vero and Rousettus cell lines, and summarize newly discovered properties of the cell lines that may pertain to pathogen discovery.
Highlights
Infection with important zoonotic pathogens, including Nipah and Hendra viruses [1,2], Ebola, Marburg, and Ravn viruses [3,4,5], coronaviruses [6,7], and lyssaviruses [8,9,10] has been found in apparently healthy bats
To facilitate epidemiological and biochemical studies of viruses adapted to fruit bats we immortalized primary cells from the Egyptian rousette (Rousettus aegyptiacus) and obtained the cell lines R06E, R05T and R05R [15]
Thereafter, R06E and R05T were provided to several laboratories until we realized that the supplied R06E cultures were not from Egyptian rousette but appeared to be Vero cells, suggesting either a contamination of the distributed R06E vials or an accidental switch of cell lines
Summary
Infection with important zoonotic pathogens, including Nipah and Hendra viruses [1,2], Ebola, Marburg, and Ravn viruses [3,4,5], coronaviruses [6,7], and lyssaviruses [8,9,10] has been found in apparently healthy bats. To facilitate epidemiological and biochemical studies of viruses adapted to fruit bats we immortalized primary cells from the Egyptian rousette (Rousettus aegyptiacus) and obtained the cell lines R06E, R05T and R05R [15]. Thereafter, R06E and R05T were provided to several laboratories until we realized that the supplied R06E cultures were not from Egyptian rousette but appeared to be Vero cells, suggesting either a contamination of the distributed R06E vials or an accidental switch of cell lines. We observed unexpected properties in intentionally-spiked cultures, an ability for occluded contamination that we consider to be important due to the wide distribution of Vero cells, and present data suggesting that cell subpassage protocols and incubation temperature of infected cultures may be contributing parameters for successful isolation of viruses from field samples or recovery of recombinant viruses
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