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

• Published in: Journal of veterinary diagnostic investigation Vol. 19; no. 6; pp. 635 - 642
• Main Authors: Lopez, L, Venteo, A, Aguirre, E, Garcia, M, Rodriguez, M.J, Amusategui, I, Tesouro, M.A, Vela, C, Sainz, A, Rueda, P
• Format: Journal Article
• Language: English
• Published: United States 01.11.2007
• Subjects: animal pathogenic bacteria; Animals; Antibodies, Bacterial - blood; Antigens, Bacterial - blood; Antigens, Bacterial - genetics; bacterial antigens; Baculoviridae - genetics; calibration; Cloning, Molecular; disease diagnosis; dog diseases; Dog Diseases - blood; Dog Diseases - diagnosis; Dogs; Ehrlichia canis; Ehrlichia canis - immunology; ehrlichiosis; Ehrlichiosis - blood; Ehrlichiosis - diagnosis; Ehrlichiosis - veterinary; enzyme-linked immunosorbent assay; Enzyme-Linked Immunosorbent Assay - veterinary; Fluorescent Antibody Technique, Indirect; immunoglobulin G; new methods; pets; recombinant antigens; Recombinant Proteins; Reproducibility of Results; Sensitivity and Specificity; serodiagnosis
• ISSN: 1040-6387; 1943-4936
• DOI: 10.1177/104063870701900604

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.