Serodiagnosis of cysticercosis by ELISA-inhibition test using monoclonal antibodies

, Yong, T S; , Yeo, I S; , Seo, J H; , Chang, J K; , Lee, J S; , Kim, T S; , Jeong, G H

doi:1993.31.2.149

Korean J Parasito > Volume 31(2):1993 > Article

Original Article


Korean J Parasitol. 1993 Jun;31(2):149-156. English. Published online Mar 20, 1994. http://dx.doi.org/10.3347/kjp.1993.31.2.149
Copyright © 1993 by The Korean Society for Parasitology


Serodiagnosis of cysticercosis by ELISA-inhibition test using monoclonal antibodies
T S Yong,^*¹I S Yeo,¹J H Seo,¹J K Chang,¹J S Lee,²T S Kim,² and G H Jeong³
¹Department of Parasitology, Yonsei University College of Medicine, Seoul 120-752, Korea.

Received February 26, 1993; Accepted March 29, 1993.
Abstract
Monoclonal antibodies (Mabs) were produced against crude scolex extract of T. solium metacestodes, and applied to ELISA-inhibition test for improving the specificity of serodiagnosis of human cysticercosis. Four hybridomas secreting species-specific anticysticercal Mabs (Cya-1, Cya-7, Cya-28 and Cya-31) were selected. Each Mab reacted on antigenic components of 25.5 kDa (Cya-1), 28 kDa (Cya-7), 87.5 kDa (Cya-28), and 12.5 kDa (Cya-31). IFA showed that Cya-1 was located at the calcium corpuscles, and Cya-7 at the loose connective tissue of T. solium metacestode scolex. Cya-28 and Cya-31 reacted on the tegument of the scolex. By conventional ELISA, 23 out of 28 (82.1%) cysticercosis patients were found serologically positive, but 1 out of 9 (11.1%) sparganosis cases and 6 out of 31 (19.4%) paragonimiasis cases showed false positives. By ELISA-inhibition test using species-specific anti-cysticercal Mab Cya-7, 19 out of 28 (67.9%) cysticercosis cases were found serologically positive, but there were no false positives in other parasitic infections.

Figures


	Fig. 1 SDS-PAGE pattern of crude scolex extract of T. solium metacestodes in 5-15% linear gradient gel, stained gel, stained with Coomassie brilliant blue R-250.


	Fig. 2 Findings of immunoblots reacted on species-specific anti-cysticercal Mabs: 1) Cya-1 (25.5 kDa), 2) Cya-7 (28 kDa), 3) Cya-28 (87.5 kDa), 4) Cya-31 (12.5 kDa), 5) immune mouse serum and 6) the protein band pattern of crude scolex extract of T. solium metacestodes on a nitrocellulose paper stained with Amido-Black B.

Fig. 3
Localization of Mabs by indirect fluorescent antibody technique. Frozen sections of scolices of T. solium metacestode were reacted on: (A) Cya-1 showing positive reactions at calcium corpuscles (×100), (B) a magnification (×200) of Fig. 1, (C) Cya-7 showing positive reactions at the loose connective tissue of a scolex (×100), (D) Cya-31 showing positive reactions at the tegument of the parasite (×100), and (E) immune mouse serum as a positive control. (▾: calcium corpuscle, C: loose connective tissue, T: tegument).

Fig. 4
Distribution of absorbance values of human sera in cysticercosis (Cy), sparganosis (Sp), paragonimiasis (Pw) cases and normal controls (NHS) against crude scolex extract of T. solium metacestodes by conventional ELISA. Twenty-three out of 28 (82.1%) cysticercosis patients were found pasitive, but 1 out of 9 (11.1%) sparganosis cases and 6 out of 31 (19.4%) paragonimiasis cases showed false positive.


	Fig. 5 Inhibition rate (%) of Mab by human sera in the ELISA-inhibition test using species-specific anti-cysticercal Mab (Cya-7) directed against an exposing antigen in infection for serodiagnosis of cysticercosis. The cut-off value is set at 12%. Nineteen out of 28 (67.9%) cysticercosis cases were found positive, and there were no false positives in other infections.

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