title: Production of a Sindbis/Eastern Equine Encephalitis chimeric virus inactivated cell culture antigen.
abstract: Eastern Equine Encephalitis virus (EEEV) is a medically important pathogen that can cause severe encephalitis in humans, with mortality rates ranging from 30 to 80%. Unfortunately there are no antivirals or licensed vaccines available for human use, and laboratory diagnosis is essential to differentiate EEEV infection 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 EEEV immunoglobulin M antibody-capture enzyme-linked immunosorbent assay (MAC-ELISA). However, EEEV is classified as a HHS select agent and requires biosafety level (BSL) three containment, limiting EEEV antigen production in non-select agent and BSL-2 laboratories. A recombinant Sindbis virus (SINV)/EEEV has been constructed for use under BSL-2 conditions and is not regulated as a select agent. Cell culture production of inactivated EEEV antigen from SINV/EEEV for use in the EEEV MAC-ELISA is reported here. Cell culture conditions and inactivation procedures were analyzed for SINV/EEEV using a recently developed antigen production algorithm, with the MAC-ELISA as the performance indicator.
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: 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: D000818:Animals; D000914:Antibodies, Viral / Q000097:blood; D000956:Antigens, Viral / Q000235:genetics* / Q000276:immunology / Q000302:isolation & purification*; D004663:Encephalitis Virus, Eastern Equine / Q000235:genetics*; D004683:Encephalomyelitis, Equine / Q000175:diagnosis*; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods; D006736:Horses; D006801:Humans; D007075:Immunoglobulin M / Q000097:blood; D012845:Sindbis Virus / Q000235:genetics* / Q000254:growth & development; D014776:Virus Cultivation / Q000379:methods
chemical_list: D000914:Antibodies, Viral; D000956:Antigens, Viral; D007075:Immunoglobulin M
keywords: Antigen; Beta-propiolactone; Gamma-irradiation; Immunoglobulin M antibody-capture enzyme-linked immunosorbent assay (MAC-ELISA); Sindbis/Eastern Equine Encephalitis chimeric virus