Immunological differentiation of root-knot nematodes (Meloidogyne spp) using monoclonal and polyclonal antibodies

Abstract

Polyclonal and monoclonal antibodies were produced to supernatants of soluble proteins from homogenates of root-knot nematodes (Meloidogyne spp.). The _antibodies were used to investigate differences between second-stage juveniles and adults of the three major root-knot species by ELISA and immunoblotting. Most antigens revealed by the polyclonal antibody were conserved between species but several quantitative differences were observed; large differences were observed between developmental stages. Cluster analysis and principal coordinate analysis using 45 monoclonal antibodies, showing partial discrimination between adult females of the nematode species, divided the nematodes into two groups; one group made up of M. arenaria and a second group composed of M. incognita and M. javanica. Three monoclonal antibodies, exhibiting partial discrimination in the order of producing coefficients of variation between the three species of nematode in excess of 40%, were selected as candidates for a diagnostic assay and immunoblotting analysis. An ELISA based assay using the three monoclonal antibodies in concomitant assays against adult females was able to distinguish the three principal species of root-knot nematodes; the same assay was unable to discriminate between the different species using second-stage juveniles.