title: Development of an algorithm for production of inactivated arbovirus antigens in cell culture.
abstract: Arboviruses are medically important pathogens that cause human disease ranging from a mild fever to encephalitis. Laboratory diagnosis is essential to differentiate arbovirus infections from other pathogens with similar clinical manifestations. The Arboviral Diseases Branch (ADB) reference laboratory at the CDC Division of Vector-Borne Diseases (DVBD) produces reference antigens used in serological assays such as the virus-specific immunoglobulin M antibody-capture enzyme-linked immunosorbent assay (MAC-ELISA). Antigen production in cell culture has largely replaced the use of suckling mice; however, the methods are not directly transferable. The development of a cell culture antigen production algorithm for nine arboviruses from the three main arbovirus families, Flaviviridae, Togaviridae, and Bunyaviridae, is described here. Virus cell culture growth and harvest conditions were optimized, inactivation methods were evaluated, and concentration procedures were compared for each virus. Antigen performance was evaluated by the MAC-ELISA at each step of the procedure. The antigen production algorithm is a framework for standardization of methodology and quality control; however, a single antigen production protocol was not applicable to all arboviruses and needed to be optimized for each virus.
authors.affiliation: Arboviral Diseases Branch, Division of Vector-borne Diseases, Centers for Disease Control and Prevention, 3156 Rampart Road, Fort Collins, CO 80521, USA. Electronic address: bvv0@cdc.gov.
authors.forename: C H
authors.initials: CH
authors.lastname: Goodman
authors.affiliation: Arboviral Diseases Branch, Division of Vector-borne Diseases, Centers for Disease Control and Prevention, 3156 Rampart Road, Fort Collins, CO 80521, USA.
authors.forename: B J
authors.initials: BJ
authors.lastname: Russell
authors.affiliation: Arboviral Diseases Branch, Division of Vector-borne Diseases, Centers for Disease Control and Prevention, 3156 Rampart Road, Fort Collins, CO 80521, USA.
authors.forename: J O
authors.initials: JO
authors.lastname: Velez
authors.affiliation: Arboviral Diseases Branch, Division of Vector-borne Diseases, Centers for Disease Control and Prevention, 3156 Rampart Road, Fort Collins, CO 80521, USA.
authors.forename: J J
authors.initials: JJ
authors.lastname: Laven
authors.affiliation: Rickettsial Zoonosis Branch, Division of Vector Borne Diseases, CDC, 1600 Clifton Road, Atlanta, GA 30333, USA.
authors.forename: W L
authors.initials: WL
authors.lastname: Nicholson
authors.affiliation: Division of Scientific Resources, CDC, 1600 Clifton Road, Atlanta, GA 30333, USA.
authors.forename: D A
authors.initials: DA
authors.lastname: Bagarozzi
authors.affiliation: Division of Scientific Resources, CDC, 1600 Clifton Road, Atlanta, GA 30333, USA.
authors.forename: J L
authors.initials: JL
authors.lastname: Moon
authors.affiliation: Division of Scientific Resources, CDC, 1600 Clifton Road, Atlanta, GA 30333, USA.
authors.forename: K
authors.initials: K
authors.lastname: Bedi
authors.affiliation: Arboviral Diseases Branch, Division of Vector-borne Diseases, Centers for Disease Control and Prevention, 3156 Rampart Road, Fort Collins, CO 80521, USA.
authors.forename: B W
authors.initials: BW
authors.lastname: Johnson
mesh_terms: D000465:Algorithms; D000818:Animals; D000956:Antigens, Viral / Q000302:isolation & purification*; D002043:Bunyaviridae / Q000737:chemistry / Q000254:growth & development* / Q000502:physiology; D018929:Cell Culture Techniques; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods; D018067:Flaviviridae / Q000737:chemistry / Q000254:growth & development* / Q000502:physiology; D006801:Humans; D012015:Reference Standards; D014035:Togaviridae / Q000737:chemistry / Q000254:growth & development* / Q000502:physiology; D014776:Virus Cultivation / Q000379:methods; D038301:Virus Inactivation
chemical_list: D000956:Antigens, Viral
keywords: Antigen; Arbovirus; Beta-propiolactone; Gamma-irradiation; Immunoglobulin M antibody-capture enzyme-linked immunosorbent assay (MAC-ELISA)
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