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Research Project: Countermeasures to Control and Eradicate Foreign Animal Diseases of Swine

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Title: Detection and quantification of African swine fever in MA104 cells

Author
item RAI, AYUSHI - Oak Ridge Institute For Science And Education (ORISE)
item Pruitt, Sarah
item RAMIEREZ-MEDINA, ELIZABETH - University Of Connecticut
item VUONO, ELIZABETH - University Of Mississippi
item SILVA, EDIANE - University Of Kansas
item VELAZQUEZ-SALINAS, LAURO - University Of Kansas
item CARRILLO, CONSUELO - Animal And Plant Health Inspection Service (APHIS)
item Gladue, Douglas
item Borca, Manuel

Submitted to: Bio-protocol
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 4/2/2020
Publication Date: 4/20/2020
Citation: Rai, A., Pruitt, S.E., Ramierez-Medina, E., Vuono, E., Silva, E., Velazquez-Salinas, L., Carrillo, C., Gladue, D.P., Borca, M.V. 2020. Detection and quantification of African swine fever in MA104 cells. Bio-protocol. https://doi.org/10.21769/BioProtoc.3955.
DOI: https://doi.org/10.21769/BioProtoc.3955

Interpretive Summary: African swine fever virus (ASFV) causes a devastating disease in swine, called African swine fever (ASF), that is currently spreading across Europe and Asia. A cell line is required to diagnose this disease. Currently there is no cell line for this purpose, and only labs with access to primary swine macrophages can perform this function. Here we provide the protocol in detail on how to detect and quantify ASFV in Ma-104 cells. This protocol will be useful to all diagnostic labs worldwide.

Technical Abstract: Detection of live African swine fever virus(ASFV) has historically relied on the use of primary swine macrophages (PSM). PSM do not replicate and have to be isolated fresh from donor swine. We previously identified that a MA-104 (ATCC# CRL-2378.1), a commercially available cell line isolated from African green monkey cells (Cercopithecus aethiops) kidney epithelial cells, supports the detection of ASFV from field samples with a sensitivity comparable to that of primary swine macrophages. MA104 could thus be used as substitute for primary swine macrophages to save significant lead time by avoiding the production of primary swine macrophages. Which require collection of swine blood or lungs, which is often not readily available in most veterinary diagnostic laboratories.