title: Methods for growing and titrating African swine fever virus: field and laboratory samples.
abstract: Growing African swine fever virus (ASFV) isolates obtained mainly from the field, but also engineered in the laboratory, is a critical step for diagnosis, titration, or virus infection studies. This unit describes a set of methods and protocols to produce and titrate any ASFV strain in cell cultures. The procedures include (1) basic techniques to prepare virus-sensitive target cells; (2) strategies for growth, concentration, and purification of virus stocks; and (3) the semi-quantitative (end dilution) and quantitative (plaque) assays for the determination of viral titers, and the use of different ASFV-sensitive cells as targets for virus production and titration.
authors.affiliation: Centro de Biologia Molecular "Severo Ochoa" (CSIC-UAM), Universidad Autonoma de Madrid, Madrid, Spain.
authors.forename: Angel L
authors.initials: AL
authors.lastname: Carrascosa
authors.affiliation: Centro de Biologia Molecular "Severo Ochoa" (CSIC-UAM), Universidad Autonoma de Madrid, Madrid, Spain.
authors.forename: M Jose
authors.initials: MJ
authors.lastname: Bustos
authors.affiliation: Centro de Biologia Molecular "Severo Ochoa" (CSIC-UAM), Universidad Autonoma de Madrid, Madrid, Spain.
authors.forename: Patricia
authors.initials: P
authors.lastname: de Leon
mesh_terms: D000357:African Swine Fever / Q000821:virology*; D000358:African Swine Fever Virus / Q000254:growth & development* / Q000302:isolation & purification* / Q000502:physiology; D000818:Animals; D018929:Cell Culture Techniques / Q000379:methods*; D002460:Cell Line; D002478:Cells, Cultured; D002499:Centrifugation, Density Gradient / Q000379:methods*; D003588:Cytopathogenic Effect, Viral; D008264:Macrophages / Q000821:virology; D009000:Monocytes / Q000821:virology; D013552:Swine; D010948:Viral Plaque Assay / Q000379:methods*; D014776:Virus Cultivation / Q000379:methods*