Skip to main content
. 2021 Feb 10;15(2):e0009027. doi: 10.1371/journal.pntd.0009027

Fig 1. Experimental workflow for the identification of novel Onchocerca lupi antigens.

Fig 1

Five steps were performed: 1) Parasites (adults and mfs) and sera collection; 2) Immunoproteomic approach. Adult female parasite was lysed and proteins extracted before separating them in a SDS-PAGE were blotted into a PVDF membrane and probed using sera collected from dogs infected with O. lupi. Bands at approximately 100 and 200kDa molecular weight were identified and analysed by LC-MS/MS. Major antigen and paramyosin proteins were identified. 3) Identification of Ol-Mja nucleotide sequence. cDNA of Major antigen protein gene from different life stages of parasite were obtained by RT-PCR and analysed by bioinformatics tools. 4) Immunogenic linear epitope scanning of Ol-MJA and Ol-PARA proteins was performed by high density-peptide array screening using sera collected from uninfected dog, O. lupi-infected dog and dogs infected with other helminths. 5) Epitopes validation was performed by incubation of epitopes with serum samples of O. lupi, Dirofilaria repens, Cercopithifilaria bainae, Dirofilaria immitis and Acanthocheilonema reconditum positive dogs and serum from negative dog.