title: Multiplexed molecular assay for rapid exclusion of foot-and-mouth disease.
abstract: A nucleic acid-based multiplexed assay was developed that combines detection of foot-and-mouth disease virus (FMDV) with rule-out assays for two other foreign animal diseases and four domestic animal diseases that cause vesicular or ulcerative lesions indistinguishable from FMDV infection in cattle, sheep and swine. The FMDV "look-alike" diagnostic assay panel contains 5 PCR and 12 reverse transcriptase PCR (RT-PCR) signatures for a total of 17 simultaneous PCR amplifications for 7 diseases plus incorporating 4 internal assay controls. It was developed and optimized to amplify both DNA and RNA viruses simultaneously in a single tube and employs Luminex liquid array technology. Assay development including selection of appropriate controls, a comparison of signature performance in single and multiplex testing against target nucleic acids, as well of limits of detection for each of the individual signatures is presented. While this assay is a prototype and by no means a comprehensive test for FMDV "look-alike" viruses, an assay of this type is envisioned to have benefit to a laboratory network in routine surveillance and possibly for post-outbreak proof of freedom from foot-and-mouth disease.
authors.affiliation: Lawrence Livermore National Laboratory, 7000 East Avenue, L-371, P.O. Box 808, Livermore, CA 94550, United States.
authors.forename: Raymond J
authors.initials: RJ
authors.lastname: Lenhoff
authors.forename: Pejman
authors.initials: P
authors.lastname: Naraghi-Arani
authors.forename: James B
authors.initials: JB
authors.lastname: Thissen
authors.forename: Jason
authors.initials: J
authors.lastname: Olivas
authors.forename: A Celena
authors.initials: AC
authors.lastname: Carillo
authors.forename: Corey
authors.initials: C
authors.lastname: Chinn
authors.forename: Max
authors.initials: M
authors.lastname: Rasmussen
authors.forename: Sharon M
authors.initials: SM
authors.lastname: Messenger
authors.forename: Lynn D
authors.initials: LD
authors.lastname: Suer
authors.forename: Sally M
authors.initials: SM
authors.lastname: Smith
authors.forename: Lance F Bentley
authors.initials: LF
authors.lastname: Tammero
authors.forename: Elizabeth A
authors.initials: EA
authors.lastname: Vitalis
authors.forename: Thomas R
authors.initials: TR
authors.lastname: Slezak
authors.forename: Pamela J
authors.initials: PJ
authors.lastname: Hullinger
authors.forename: Benjamin J
authors.initials: BJ
authors.lastname: Hindson
authors.forename: Sharon K
authors.initials: SK
authors.lastname: Hietala
authors.forename: Beate M
authors.initials: BM
authors.lastname: Crossley
authors.forename: Mary T
authors.initials: MT
authors.lastname: McBride
mesh_terms: D000818:Animals; D002417:Cattle; D002418:Cattle Diseases / Q000821:virology*; D017931:DNA Primers; D005536:Foot-and-Mouth Disease / Q000175:diagnosis*; D029544:Foot-and-Mouth Disease Virus / Q000235:genetics / Q000302:isolation & purification; D016133:Polymerase Chain Reaction / Q000379:methods* / Q000592:standards; D012015:Reference Standards; D012680:Sensitivity and Specificity; D012756:Sheep; D012757:Sheep Diseases / Q000821:virology*; D013552:Swine; D013553:Swine Diseases / Q000821:virology*
chemical_list: D017931:DNA Primers