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Journal of Clinical Microbiology, April 2000, p. 1409-1413, Vol. 38, No. 4
0095-1137/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.

Development of a Highly Specific Recombinant Toxocara canis Second-Stage Larva Excretory-Secretory Antigen for Immunodiagnosis of Human Toxocariasis

Hiroshi Yamasaki,1,* Kunioki Araki,2 Patricia Kim Chooi Lim,3 Ngah Zasmy,4 Joon Wah Mak,5 Radzan Taib,3 and Takashi Aoki1

Department of Parasitology, Juntendo University School of Medicine,1 and Department of Microbiology, National Institute of Public Health,2 Tokyo, Japan, and Division of Molecular Pathology3 and Director's Laboratory,4 Institute for Medical Research, Kuala Lumpur, and Faculty of Medicine and Health Science, Universiti Putra Malaysia, Selangor,5 Malaysia

Received 10 September 1999/Returned for modification 26 October 1999/Accepted 3 January 2000

The specificity of the recombinant Toxocara canis antigen developed for the immunodiagnosis of human toxocariasis was compared with that of the excretory-secretory antigen from T. canis second-stage larvae (TES) by enzyme-linked immunosorbent assay. A total of 153 human serum samples from patients infected with 20 different helminths, including 11 cases of toxocariasis, were examined. No false-negative reactions were observed for the toxocariasis cases. When the TES was used at concentrations of 0.5 and 0.125 µg/ml, cross-reactions were observed in 79 (55.6%) and 61 (43.0%) of 142 cases, respectively. In contrast, when the recombinant antigen was tested at a concentration of 0.5 µg/ml, cross-reactions were observed in 19 (13.4%) of 142 cases. At a concentration of 0.125 µg/ml, however, the cross-reaction rate decreased sharply to only 2.1%, corresponding to 3 of 142 cases. The cross-reactions occurred with one case each of gnathostomiasis, paragonimiasis with Paragonimus miyazakii, and spirometriasis, in which high antibody titers were detected. In addition, the recombinant antigen showed negative reactions with serum samples from patients infected with Ascaris and hookworms, which are the most common parasites in the world. These findings are also supported by experiments with animals infected with Ascaris and hookworm. From these results, the recombinant antigen is highly specific for toxocariasis and may provide more reliable diagnostic results than other methods.

* Corresponding author. Mailing address: Department of Parasitology, Juntendo University School of Medicine, 2-1-1 Hongo, Bunkyo-ku, Tokyo 113-0033, Japan. Phone: 81-3-5802-1043. Fax: 81-3-5800-0476. E-mail: hyamasak{at}med.juntendo.ac.jp.

Journal of Clinical Microbiology, April 2000, p. 1409-1413, Vol. 38, No. 4
0095-1137/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.


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