title: Initial development and preliminary evaluation of a multiplex bead assay to detect antibodies to Ehrlichia canis, Anaplasma platys, and Ehrlichia chaffeensis outer membrane peptides in naturally infected dogs from Grenada, West Indies.
abstract: Tick-borne bacteria, Ehrlichia canis, Anaplasma platys, and Ehrlichia chaffeensis are significant pathogens of dogs worldwide, and coinfections of E. canis and A. platys are common in dogs on the Caribbean islands. We developed and evaluated the performance of a multiplex bead-based assay to detect antibodies to E. canis, A. platys, and E. chaffeensis peptides in dogs from Grenada, West Indies, where E. canis and A. platys infections are endemic. Peptides from outer membrane proteins of P30 of E. canis, OMP-1X of A. platys, and P28-19/P28-14 of E. chaffeensis were coupled to magnetic beads. The multiplex peptide assay detected antibodies in dogs experimentally infected with E. canis and E. chaffeensis, but not in an A. platys experimentally infected dog. In contrast, the multiplex assay and an in-house enzyme-linked immunosorbent assay (ELISA) detected A. platys antibodies in naturally infected Grenadian dogs. Following testing of 104 Grenadian canine samples, multiplex assay results had good agreement with commercially available ELISA and immunofluorescent assay for E. canis antibody-positive dogs ( K values of 0.73 and 0.84), whereas A. platys multiplex results had poor agreement with these commercial assays ( K values of -0.02 and 0.01). Prevalence of seropositive E. canis and A. platys Grenadian dogs detected by the multiplex and commercial antibody assays were similar to previous reports. Although the multiplex peptide assay performed well in detecting the seropositive status of dogs to E. canis and had good agreement with commercial assays, better antigen targets are necessary for the antibody detection of A. platys.
authors.affiliation: Department of Diagnostic Medicine/Pathobiology and Kansas State Veterinary Diagnostic Laboratory, Kansas State University, Manhattan, KS (Wilkerson, Black, George, Nair, Ganta).|Small Animal Medicine and Surgery Department (Lanza-Perea), School of Veterinary Medicine, St. George's University, Grenada, West Indies.|Department of Pathobiology (Sharma, Gibson, Stone), School of Veterinary Medicine, St. George's University, Grenada, West Indies.
authors.forename: Melinda J
authors.initials: MJ
authors.lastname: Wilkerson
authors.affiliation: Department of Diagnostic Medicine/Pathobiology and Kansas State Veterinary Diagnostic Laboratory, Kansas State University, Manhattan, KS (Wilkerson, Black, George, Nair, Ganta).|Small Animal Medicine and Surgery Department (Lanza-Perea), School of Veterinary Medicine, St. George's University, Grenada, West Indies.|Department of Pathobiology (Sharma, Gibson, Stone), School of Veterinary Medicine, St. George's University, Grenada, West Indies.
authors.forename: Kelley E
authors.initials: KE
authors.lastname: Black
authors.affiliation: Department of Diagnostic Medicine/Pathobiology and Kansas State Veterinary Diagnostic Laboratory, Kansas State University, Manhattan, KS (Wilkerson, Black, George, Nair, Ganta).|Small Animal Medicine and Surgery Department (Lanza-Perea), School of Veterinary Medicine, St. George's University, Grenada, West Indies.|Department of Pathobiology (Sharma, Gibson, Stone), School of Veterinary Medicine, St. George's University, Grenada, West Indies.
authors.forename: Marta
authors.initials: M
authors.lastname: Lanza-Perea
authors.affiliation: Department of Diagnostic Medicine/Pathobiology and Kansas State Veterinary Diagnostic Laboratory, Kansas State University, Manhattan, KS (Wilkerson, Black, George, Nair, Ganta).|Small Animal Medicine and Surgery Department (Lanza-Perea), School of Veterinary Medicine, St. George's University, Grenada, West Indies.|Department of Pathobiology (Sharma, Gibson, Stone), School of Veterinary Medicine, St. George's University, Grenada, West Indies.
authors.forename: Bhumika
authors.initials: B
authors.lastname: Sharma
authors.affiliation: Department of Diagnostic Medicine/Pathobiology and Kansas State Veterinary Diagnostic Laboratory, Kansas State University, Manhattan, KS (Wilkerson, Black, George, Nair, Ganta).|Small Animal Medicine and Surgery Department (Lanza-Perea), School of Veterinary Medicine, St. George's University, Grenada, West Indies.|Department of Pathobiology (Sharma, Gibson, Stone), School of Veterinary Medicine, St. George's University, Grenada, West Indies.
authors.forename: Kathryn
authors.initials: K
authors.lastname: Gibson
authors.affiliation: Department of Diagnostic Medicine/Pathobiology and Kansas State Veterinary Diagnostic Laboratory, Kansas State University, Manhattan, KS (Wilkerson, Black, George, Nair, Ganta).|Small Animal Medicine and Surgery Department (Lanza-Perea), School of Veterinary Medicine, St. George's University, Grenada, West Indies.|Department of Pathobiology (Sharma, Gibson, Stone), School of Veterinary Medicine, St. George's University, Grenada, West Indies.
authors.forename: Diana M
authors.initials: DM
authors.lastname: Stone
authors.affiliation: Department of Diagnostic Medicine/Pathobiology and Kansas State Veterinary Diagnostic Laboratory, Kansas State University, Manhattan, KS (Wilkerson, Black, George, Nair, Ganta).|Small Animal Medicine and Surgery Department (Lanza-Perea), School of Veterinary Medicine, St. George's University, Grenada, West Indies.|Department of Pathobiology (Sharma, Gibson, Stone), School of Veterinary Medicine, St. George's University, Grenada, West Indies.
authors.forename: Anushka
authors.initials: A
authors.lastname: George
authors.affiliation: Department of Diagnostic Medicine/Pathobiology and Kansas State Veterinary Diagnostic Laboratory, Kansas State University, Manhattan, KS (Wilkerson, Black, George, Nair, Ganta).|Small Animal Medicine and Surgery Department (Lanza-Perea), School of Veterinary Medicine, St. George's University, Grenada, West Indies.|Department of Pathobiology (Sharma, Gibson, Stone), School of Veterinary Medicine, St. George's University, Grenada, West Indies.
authors.forename: Arathy D S
authors.initials: AD
authors.lastname: Nair
authors.affiliation: Department of Diagnostic Medicine/Pathobiology and Kansas State Veterinary Diagnostic Laboratory, Kansas State University, Manhattan, KS (Wilkerson, Black, George, Nair, Ganta).|Small Animal Medicine and Surgery Department (Lanza-Perea), School of Veterinary Medicine, St. George's University, Grenada, West Indies.|Department of Pathobiology (Sharma, Gibson, Stone), School of Veterinary Medicine, St. George's University, Grenada, West Indies.
authors.forename: Roman R
authors.initials: RR
authors.lastname: Ganta
mesh_terms: D000709:Anaplasma / Q000276:immunology / Q000302:isolation & purification; D000818:Animals; D000907:Antibodies, Bacterial / Q000097:blood; D004283:Dog Diseases / Q000097:blood / Q000175:diagnosis* / Q000382:microbiology; D004285:Dogs; D041061:Ehrlichia canis / Q000276:immunology / Q000302:isolation & purification; D018469:Ehrlichia chaffeensis / Q000276:immunology / Q000302:isolation & purification; D016873:Ehrlichiosis / Q000175:diagnosis / Q000662:veterinary*; D004797:Enzyme-Linked Immunosorbent Assay / Q000662:veterinary; D019425:Grenada; D011237:Predictive Value of Tests
chemical_list: D000907:Antibodies, Bacterial
keywords: Anaplasma; Ehrlichia; canine antibodies; multiplex bead assay; peptides