title: Laboratory validation of a lateral flow device for the detection of CyHV-3 antigens in gill swabs.
abstract: Cyprinid herpesvirus-3 (CyHV-3) induces the highly contagious koi herpesvirus disease (KHVD) and may result in significant economic losses to the ornamental and food-producing carp industry. Suspicion of KHVD is triggered by clinical signs and confirmed using laboratory techniques. The latter are labour- and time-consuming, require specialised equipment and trained personnel. For rapid, on-site detection of CyHV-3, a lateral flow device (LFD) was developed using two monoclonal antibodies directed towards the viral glycoprotein ORF65. The LFD was highly specific with analytical and diagnostic specificities of 100%. Analytical sensitivity ranged between 1.25×10(2) and 2.40×10(4) plaque forming units per ml for isolates originating from geographically distinct regions. In experimentally infected carp, CyHV-3 was detected as early as 4-5 days post infection. Diagnostic sensitivities of 52.6% and 72.2% relative to PCR were recorded, depending on the viral isolate used. When onset of mortality was taken as reference, diagnostic sensitivities increased to 67.0% and 93.3%. The diagnostic sensitivity for freshly found-dead animals was 100%, irrespective of the virus isolate used. Given the high specificity and ease-of-use for on-site detection of CyHV-3, the LFD was regarded fit for purpose as a first-line diagnostic tool for the identification of acute CyHV-3 infections in KHVD affected (koi) carp.
authors.affiliation: Okapi Sciences NV, B-3001 Heverlee, Belgium. Electronic address: Robert.vrancken@okapi-sciences.com.
authors.forename: R
authors.initials: R
authors.lastname: Vrancken
authors.forename: M
authors.initials: M
authors.lastname: Boutier
authors.forename: M
authors.initials: M
authors.lastname: Ronsmans
authors.forename: A
authors.initials: A
authors.lastname: Reschner
authors.forename: T
authors.initials: T
authors.lastname: Leclipteux
authors.forename: F
authors.initials: F
authors.lastname: Lieffrig
authors.forename: A
authors.initials: A
authors.lastname: Collard
authors.forename: C
authors.initials: C
authors.lastname: Mélard
authors.forename: S
authors.initials: S
authors.lastname: Wera
authors.forename: J
authors.initials: J
authors.lastname: Neyts
authors.forename: N
authors.initials: N
authors.lastname: Goris
authors.forename: A
authors.initials: A
authors.lastname: Vanderplasschen
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal; D000914:Antibodies, Viral; D000956:Antigens, Viral / Q000032:analysis*; D002347:Carps; D005393:Fish Diseases / Q000175:diagnosis* / Q000821:virology; D005880:Gills / Q000821:virology*; D006564:Herpesviridae / Q000276:immunology / Q000302:isolation & purification*; D006566:Herpesviridae Infections / Q000175:diagnosis / Q000662:veterinary* / Q000821:virology; D007118:Immunoassay / Q000295:instrumentation / Q000379:methods; D012680:Sensitivity and Specificity; D014730:Veterinary Medicine / Q000295:instrumentation* / Q000379:methods; D014773:Virology / Q000295:instrumentation* / Q000379:methods
chemical_list: D000911:Antibodies, Monoclonal; D000914:Antibodies, Viral; D000956:Antigens, Viral
keywords: Cyprinid herpesvirus-3; Immunochromatographic assay; Koi herpesvirus disease; Lateral flow device