title: Development of a sensitive and specific indirect enzyme-linked immunosorbent assay based on a baculovirus recombinant antigen for detection of specific antibodies against Ehrlichia canis.
abstract: An indirect enzyme-linked immunosorbent assay (ELISA) based on baculovirus recombinant P30 protein of Ehrlichia canis and the 1BH4 anticanine IgG monoclonal antibody was developed and evaluated by examining a panel of 98 positive and 157 negative sera using the indirect fluorescent antibody (IFA) test as the reference technique. The P30-based ELISA appeared to be sensitive and specific (77.55% and 95.54%, respectively) when qualitative results (positive/negative) were compared with those of the IFA test; the coefficient of correlation (R) between the 2 tests was 0.833. Furthermore, it was possible to establish a mathematical formula for use in comparing the results of both techniques. These results indicate that recombinant P30 antigen-based ELISA is a suitable alternative of the IFA test for simple, consistent, and rapid serodiagnosis of canine ehrlichiosis. Moreover, the use of this recombinant protein as antigen offers a great advantage for antigen preparation in comparison with other techniques in which the whole E. canis organism is used as antigen.
authors.affiliation: Inmunología y Genética Aplicada S.A. (INGENASA), Hnos. García Noblejas 41, 28037 Madrid, Spain.
authors.forename: Lissett
authors.initials: L
authors.lastname: López
authors.forename: Angel
authors.initials: A
authors.lastname: Venteo
authors.forename: Enara
authors.initials: E
authors.lastname: Aguirre
authors.forename: Marga
authors.initials: M
authors.lastname: García
authors.forename: Majosé
authors.initials: M
authors.lastname: Rodríguez
authors.forename: Inmaculada
authors.initials: I
authors.lastname: Amusátegui
authors.forename: Miguel A
authors.initials: MA
authors.lastname: Tesouro
authors.forename: Carmen
authors.initials: C
authors.lastname: Vela
authors.forename: Angel
authors.initials: A
authors.lastname: Sainz
authors.forename: Paloma
authors.initials: P
authors.lastname: Rueda
mesh_terms: D000818:Animals; D000907:Antibodies, Bacterial / Q000097:blood*; D000942:Antigens, Bacterial / Q000097:blood* / Q000235:genetics; D016367:Baculoviridae / Q000235:genetics; D003001:Cloning, Molecular; D004283:Dog Diseases / Q000097:blood / Q000175:diagnosis*; D004285:Dogs; D041061:Ehrlichia canis / Q000276:immunology*; D016873:Ehrlichiosis / Q000097:blood / Q000175:diagnosis / Q000662:veterinary*; D004797:Enzyme-Linked Immunosorbent Assay / Q000662:veterinary*; D019084:Fluorescent Antibody Technique, Indirect; D011994:Recombinant Proteins; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity
chemical_list: D000907:Antibodies, Bacterial; D000942:Antigens, Bacterial; D011994:Recombinant Proteins