Author
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MECHAM, JAMES |
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Submitted to: Journal of Virological Methods
Publication Type: Peer Reviewed Journal Publication Acceptance Date: 3/6/2006 Publication Date: 8/1/2006 Citation: Mecham, J.O. 2006. Detection and titration of bluetongue virus in culicoides insect cell culture by an antigen-capture enzyme-linked immunosorbent assay. Journal of Virological Methods. 135:269-271 Interpretive Summary: Bluetongue virus (BTV) infects sheep, cattle and other ruminants and is transmitted by Culicoides spp. of biting midges. Inoculation of susceptible sheep, chicken eggs and mammalian cell culture has been used to isolate and quantify virus from biological samples. In this laboratory, cell lines have been produced from C. sonorensis that have proven to be particularly useful for various studies on BTV and the insect vector. However, since BTV does not produce detectable damage in these cells, indirect methods, such as co-cultivation with susceptible mammalian cells, are necessary for detection and quantifying virus replication in the insect cells. This paper describes the application of an antigen-capture enzyme-linked immunosorbent assay (Ag-cap ELISA) for direct detection and quantification of BTV in one of these insect cell lines. This assay was comparable in sensitivity to detection and quantification of virus in susceptible vertebrate cell culture using a conventional assay system. It will facilitate the use of the Culicoides cell lines in research questions and for isolation and quantification of BTV from biological samples. Technical Abstract: Bluetongue virus (BTV) infects sheep, cattle and other ruminants and is transmitted by Culicoides spp. of biting midges. Virus is typically isolated and characterized by infection of susceptible vertebrate cells that undergo detectable and measurable cytopathic effects. Cell lines derived from C. sonorensis may be useful for virus isolation and studies to better understand BTV replication in the insect vector. However, their use is hampered because BTV infection does not produce significant cytopathic effects in these insect cell cultures. Detection of virus replication in these cells typically requires co-cultivation with susceptible vertebrate cell culture. This report describes the use of an antigen-capture enzyme-linked immunosorbent assay (Ag-Cap ELISA) for direct detection and titration of BTV in cultures of a Culicoides cell line. This assay should facilitate use of this cell line for virus isolation, titration and studies of BTV replication. |
