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Journal of Veterinary Diagnostic Investigation Vol. 19 Issue 6, 635-642
Copyright © 2007 by the American Association of Veterinary Laboratory Diagnosticians
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Full Scientific Reports

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

Lissett López, Angel Venteo, Enara Aguirre, Marga García, MaJosé Rodríguez, Inmaculada Amusátegui, Miguel A. Tesouro, Carmen Vela, Angel Sainz and Paloma Rueda1

Correspondence: 1Corresponding Author: Paloma Rueda, Inmunología y Genética Aplicada S.A. (INGENASA), Hnos. García Noblejas 41, 28037 Madrid, Spain, e-mail: prueda{at}ingenasa.es

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.

Key Words: Ehrlichia canis • indirect enzyme-linked immunosorbent assay (ELISA) • indirect fluorescent antibody (IFA) • P30 recombinant protein • serological detection






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