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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

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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				L. Lopez, A. Venteo, M. Garcia, A. Camunas, A. Ranz, J. Garcia, J. Sarraseca, C. Anaya, and P. Rueda Antigen-capture blocking enzyme-linked immunosorbent assay based on a baculovirus recombinant antigen to differentiate Transmissible gastroenteritis virus from Porcine respiratory coronavirus antibodies J Vet Diagn Invest, September 1, 2009; 21(5): 598 - 608. [Abstract] [Full Text] [PDF]