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

J Vet Diagn Invest. 2007 Nov;19(6):635-42. doi: 10.1177/104063870701900604.

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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Antibodies, Bacterial / blood*
  • Antigens, Bacterial / blood*
  • Antigens, Bacterial / genetics
  • Baculoviridae / genetics
  • Cloning, Molecular
  • Dog Diseases / blood
  • Dog Diseases / diagnosis*
  • Dogs
  • Ehrlichia canis / immunology*
  • Ehrlichiosis / blood
  • Ehrlichiosis / diagnosis
  • Ehrlichiosis / veterinary*
  • Enzyme-Linked Immunosorbent Assay / veterinary*
  • Fluorescent Antibody Technique, Indirect
  • Recombinant Proteins
  • Reproducibility of Results
  • Sensitivity and Specificity

Substances

  • Antibodies, Bacterial
  • Antigens, Bacterial
  • Recombinant Proteins