Preparation of a monoclonal antibody specific for Entamoeba dispar and its ability to distinguish E. dispar from E. histolytica.
A monoclonal antibody (MAb), MAb ED17 (immunoglobulin G2a [IgG2a]), prepared against trophozoites of Entamoeba dispar SAW1734RclAR cultured monoxenically with Crithidia fasciculata, reacted with 25 of 26 isolates of E. dispar by an indirect fluorescent-antibody test. In contrast, the MAb failed to react with any of 20 isolates of E. histolytica or other enteric protozoan parasites. Western blot (immunoblot) analysis showed that the molecular mass of the E. dispar antigen recognized by the MAb was 160 kDa under reduced conditions. Immunoelectron microscopy revealed that the antigen was mainly located on digested C. fasciculata, but not on undigested organisms. Double staining with a mixture of MAb ED17 and MAb 4G6 (an IgG1 MAb which reacts exclusively with E. histolytica), followed by incubation with a mixture of fluorescein isothiocyanate-labeled anti-mouse IgG2a and tetramethylrhodamine isothiocyanate-labeled anti-mouse IgG1 antibodies, simultaneously identified mixed populations of E. dispar and E. histolytica. This method may prove to be useful for the accurate identification of E. dispar and E. histolytica, even in mixed infections.
Tachibana
H
H
Department of Infectious Diseases, Tokai University School of Medicine, Isehara, Japan. htachiba@is.icc.u-tokai.ac.jp
Kobayashi
S
S
Kaneda
Y
Y
Takeuchi
T
T
Fujiwara
T
T
D000818:Animals; D000911:Antibodies, Monoclonal; D000918:Antibody Specificity; D000953:Antigens, Protozoan / Q000276:immunology; D015153:Blotting, Western; D004748:Entamoeba histolytica / Q000276:immunology* / Q000302:isolation & purification* / Q000648:ultrastructure; D004749:Entamoebiasis / Q000175:diagnosis*; D005260:Female; D005455:Fluorescent Antibody Technique / Q000379:methods; D051379:Mice; D008807:Mice, Inbred BALB C; D008854:Microscopy, Electron; D013045:Species Specificity; D013194:Staining and Labeling
D000911:Antibodies, Monoclonal; D000953:Antigens, Protozoan