title: Echinococcus multilocularis: purification and characterization of glycoprotein antigens with serodiagnostic potential for canine infection.
abstract: We show that a conventionally purified glycoprotein component of Echinococcus multilocularis protoscolex, designated as Emgp-89, may be useful as a serodiagnostic antigen for detecting E. multilocularis infection in dogs domesticated in endemic areas. Emgp-89 was obtained from the parasite material by a simple procedure using Con A-agarose and subsequent gel filtration chromatography. The purified fraction showed a molecular weight of >4000kDa upon gel filtration and reacted with a series of lectins that specifically bind to mannose, galactose, N-acetylglucosamine, and N-acetylgalactosamine. Subsequently, serodiagnostic performance of Emgp-89 was evaluated through enzyme-linked immunosorbent assays (ELISAs) by using sera from normal, domestic dogs and dogs infected with other helminths. Emgp-89 positively reacted with all 16 serum samples from E. multilocularis-infected dogs, thus showing that this antigen is highly sensitive. On the other hand, the specificity of Emgp-89-based ELISA, determined using 41 serum samples from dogs infected with other helminths, was relatively low (83%). As an attempt to improve the specificity of Emgp-89-based ELISA, we pretreated Emgp-89 with proteinase K or sodium periodate, expecting that these treatments would enable discrimination of true positives from false positives. The ELISA value increased after treatment with sodium periodate in most false-positive samples, whereas significant decreases were observed in sera from all dogs infected with E. multilocularis. Further evaluation of this antigen should be performed using sera from dogs infected with closely-related parasites, including taeniid cestodes, which are expected to prove that this serodiagnostic system is sufficiently specific for clinical and field applications.
authors.affiliation: Hokkaido Institute of Public Health, N19 W12, Kita-Ku, Sapporo, Japan.
authors.forename: Hirokazu
authors.initials: H
authors.lastname: Kouguchi
authors.forename: Jun
authors.initials: J
authors.lastname: Matsumoto
authors.forename: Kimiaki
authors.initials: K
authors.lastname: Yamano
authors.forename: Yoshinobu
authors.initials: Y
authors.lastname: Katoh
authors.forename: Yuzaburo
authors.initials: Y
authors.lastname: Oku
authors.forename: Tomohiro
authors.initials: T
authors.lastname: Suzuki
authors.forename: Kinpei
authors.initials: K
authors.lastname: Yagi
mesh_terms: D000818:Animals; D000909:Antibodies, Helminth / Q000096:biosynthesis / Q000097:blood*; D000947:Antigens, Helminth / Q000276:immunology / Q000302:isolation & purification*; D015153:Blotting, Western / Q000662:veterinary; D002850:Chromatography, Gel / Q000662:veterinary; D004283:Dog Diseases / Q000175:diagnosis* / Q000276:immunology / Q000469:parasitology; D004285:Dogs; D004443:Echinococcosis / Q000175:diagnosis / Q000276:immunology / Q000662:veterinary*; D048210:Echinococcus multilocularis / Q000276:immunology*; D004797:Enzyme-Linked Immunosorbent Assay / Q000662:veterinary; D006023:Glycoproteins / Q000276:immunology / Q000302:isolation & purification*; D008297:Male; D016205:Sigmodontinae; D013997:Time Factors
chemical_list: D000909:Antibodies, Helminth; D000947:Antigens, Helminth; D006023:Glycoproteins