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Journal of Clinical Microbiology, August 2002, p. 2760-2765, Vol. 40, No. 8
0095-1137/02/$04.00+0     DOI: 10.1128/JCM.40.8.2760-2765.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.

Alveolar Echinococcosis: Characterization of Diagnostic Antigen Em18 and Serological Evaluation of Recombinant Em18

Yasuhito Sako,1* Minoru Nakao,1 Kazuhiro Nakaya,2 Hiroshi Yamasaki,1 Bruno Gottstein,3 Marshall W. Lightowers,4 Peter M. Schantz,5 and Akira Ito1

Department of Parasitology,1 Animal Laboratory for Medical Research, Asahikawa Medical College, Asahikawa, Japan,2 Institute of Parasitology, University of Berne, Berne, Switzerland,3 Veterinary Clinical Center, University of Melbourne, Werribee, Melbourne, Victoria, Australia,4 Division of Parasitic Diseases, NCID, Centers for Disease Control and Prevention, Atlanta, Georgia5

Received 14 March 2002/ Returned for modification 23 April 2002/ Accepted 7 May 2002

The Echinococcus multilocularis protein Em18 is one of the most promising antigens for use in serodiagnosis of alveolar echinococcosis in human patients. Here we identify an antigenic relationship between Em18 and a 65-kDa immunodominant E. multilocularis surface protein previously identified as either EM10 or EmII/3. The NH2-terminal sequence of native Em18 was determined, revealing it to be a fragment of EM10. Experiments were undertaken to investigate the effect of proteinase inhibitors on the degradation of EM10 in crude extracts of E. multilocularis protoscoleces. Em18 was found to be the product of degradation of EM10 by cysteine proteinase. A recombinant Em18 (RecEm18, derived from 349K to 508K of EM10) was successfully expressed by using Escherichia coli expression system and then evaluated for use in serodiagnosis of alveolar echinococcosis. RecEm18 was recognized by 27 (87.1%) and 28 (90.3%) of 31 serum samples from clinically and/or pathologically confirmed alveolar echinococcosis patients by enzyme-linked immunosorbent assay and immunoblotting, respectively. Of 33 serum samples from cystic echinococcosis patients, 1 was recorded as having a weak positive reaction to RecEm18; however, none of the serum samples which were tested from neurocysticercosis patients (n = 10) or healthy people (n = 15) showed positive reactions. RecEm18 has the potential for use in the differential serodiagnosis of alveolar echinococcosis.

* Corresponding author. Mailing address: Department of Parasitology, Asahikawa Medical College, Asahikawa, Hokkaido 078-8510, Japan. Phone: 81-166-68-2422. Fax: 81-166-68-2429. E-mail: yasusako{at}asahikawa-med.ac.jp.

Journal of Clinical Microbiology, August 2002, p. 2760-2765, Vol. 40, No. 8
0095-1137/02/$04.00+0     DOI: 10.1128/JCM.40.8.2760-2765.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.



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