Analysis Of Protein Patterns Of Gnathostoma Spinigerum Larvae In Swamp Eeels (Monopterus Alba) By Immunoblotting

Abstract

Gnathostoma spinigerum is an intestine nematode in carnivore but can cause human gnathostomiasis. Man is a paratenic host and can acquires the infection by eating non cooked or inadequately processed intermediate hosts, e.g. freshwater fishes, frogs, birds and snakeswhich contain encapsulated third stage larvae. Third stage larvaes (L3s) of G. spinigerum were most found in liver and muscle of swamp eels (Monopterus alba). The objective of this research was to analyze the specific antigens of excretory-secretory (ES) products of G. spinigerum L3s (ES-Ag Gs L3) against rabbit immunoglobulin G (IgG) polyclonal antibody-anti ES-Ag Gs L3s using Western blotting technique. The result of this research was hoped to be a basic for the development of immunodiagnostic and molecular diagnostic of gnathostomias. Excretory-secratory proteins isolation of G. spinigerum L3 were obtained by incubating L3 in RPMI 1640 medium which contains 100 units/ml penicillin G and 100 μg/ml streptomycin for four days at 37°C under 5% CO2 in air. The crude antigen protein of ES-L3-Gs were not purificated. Immunoglobulin G anti ES-Ag Gs L3s were obtained by immunize New Zealand strain white rabbits with ES-Ag Gs L3s. Bands with apparent molecular weights of 12.6, 18.6, 37.9, 41.7 and 49.5 kDa were identified from immunoblotting reactions between IgG-anti ES-Ag Gs L3s and ES-Ag Gs L3s. The proteins of ES from other nematode larvae with apparent molecular weights of 12.6, 18.6, and 37.9 were also recognized by IgG-anti ES-Ag Gs L3s. The specific proteins with molecular weights of 41.7 and 49.5 kDa were potential antigens for immunodiagnostic and molecular diagnostic of gnathostomiasis in swamp eels and human gnathostomiasis in Indonesia.