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 purific...Continue Reading
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