Serodiagnosis of sheeppox and goatpox using an indirect ELISA based on synthetic peptide targeting for the major antigen P32

The results indicated that the optimal concentration of coated recombinant antigen was 0.2 μg per well for a serum dilution of 1:10. The ELISA performed favorably when sera from sheep immunized experimentally were tested.This assay offers the prospect of synthetic peptide as antigens for indirect ELISA to detect SPPV and GTPV antibody in sheep and goat sera.Goat pox (GP) and sheep pox (SP) are malignant diseases of small ruminants causing heavy economic loss in the endemic countries. The diseases are endemic in India, Bangladesh, throughout the near and middle east, northern and central Africa [1,2]. The causative agents, sheep pox and goat pox viruses, belong to the genus Capripoxvirus in the family Poxviridae [3,4].Although experienced veterinarians readily diagnose these diseases in their acute forms, low virulence strains and other exanthemas in sheep, e.g., orf or scabby mouth Parapoxviridae can present problems for differential diagnosis. Laboratory confirmation has been reliant upon classical virological techniques including animal transmission, electron microscopy for identification of virus in clinical material and virus isolation in cell culture [2]. Sero-epidemiological surveillance has been difficult since the only antibody tests available have been based upon immunofluorescence and virus neutralisation tests in cell culture. These tests are difficult and time consuming and not readily available in countries that do not hold live viruses.P32, one of the structural proteins present in all the capripoxviruses, contains major immunogenic determinants. Here we selected two particular amino-acid sequence sits (residues 92-118 and 156-175, its character were identified by DNAstar Lasergene 7.1) and used this sequence for synthesis of one 27 amino-acid and one 20 amino-acid synthetic antigen. These synthesis peptides were used to develop a more convenient and cost-effective ELISA, that offers the prospect of reliable, high-throughput sero-surveillance on a floc