Development of latex agglutination test with recombinant NcSAG1 for the rapid detection of antibodies to Neospora caninum in cattle

Abstract: Neospora caninum, an apicomplexan protozoan parasite, is recognized as a major cause of abortion in cattle. Surface antigen 1 of N. caninum (NcSAG1) is an important immunodominant candidate for the development of a diagnostic reagent for neosporosis. The present study describes the development and evaluation of a latex agglutination test (LAT) with recombinant NcSAG1 (rNcSAG1) for the detection of antibodies to N. caninum in cattle. The rNcSAG1 gene was cloned in pET-28a and protein was expressed in Escherichia coli BL21 (DE3). Carboxylated latex particles were coated with rNcSAG1 and the degree of agreement between LAT and a commercial enzyme-linked immunosorbent assay (iscomELISA) was evaluated by using of 164 serum samples. Twenty-two (13.4%) and 23 (14.0%) of samples were positive for antibodies to N. caninum by LAT and ELISA respectively. Eighteen of 23 ELISA-positive samples were positive according to the LAT and a substantial agreement (κ =0.77) was found between the results of LAT and ELISA. The results indicated that the LAT with rNcSAG1 would be a rapid, simple, relatively inexpensive and suitable diagnostic test for detection of specific antibodies in N. caninum infection under field conditions. Improvement in purification of rNcSAG1 can reduce probable false positive reactions and so increase the degree of agreement between the LAT and ELISA. [Copyright &y& Elsevier]