Serodiagnosis of human paragonimiasis by ELISA-inhibition test using monoclonal antibodies

, Yong, T S; , Seo, J H; , Yeo, I S

doi:1993.31.2.141

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

Original Article


Korean J Parasitol. 1993 Jun;31(2):141-147. English. Published online Mar 20, 1994. http://dx.doi.org/10.3347/kjp.1993.31.2.141
Copyright © 1993 by The Korean Society for Parasitology


Serodiagnosis of human paragonimiasis by ELISA-inhibition test using monoclonal antibodies
T S Yong,^*J H Seo and I S Yeo
Department of Parasitology and Institute of Tropical Medicine, Yonsei University College of Medicine, Seoul 120-752, Korea.

Received February 06, 1993; Accepted March 20, 1993.
Abstract
ELISA-inhibition test using Paragonimus westermani specific monoclonal antibody (Mab) was investigated to improve the diagnostic specificity of paragonimiasis. By cell fusion, one hybridoma clone secreting anti-P. westermani specific Mab was selected (Pwa-14), which reacted on bands of 28 kDa, 42.5 kDa, 89 kDa and 120.5 kDa. IFA showed Pwa-14 was located at the vitelline follicles. By micro-ELISA, 100% of 22 paragonimiasis cases were found positive, but 5 of 40 clonorchiasis cases (12.5%), 3 of 26 cysticercosis cases (7.7%) showed false positive. None of 10 sparganosis patients or 28 normal controls reacted positively. On the other hand, by ELISA-inhibition test using a P. westermani specific Mab, 100% of paragonimiasis cases were found positive, and there were no positive in cysticercosis, sparganosis cases or normal controls, except 2 (5.0%) false-positive sera of 40 clonorchiasis cases. The ELISA-inhibition test using a Mab showed higher specificity in comparison with micro-ELISA for serodiagnosis of human paragonimiasis.

Figures


	Fig. 1 SDS-PAGE finding of P. westermani adult worm crude extract (Pw) stained with Coomassie blue R-250.


	Fig. 2 Immunoblots reacted on: 1) Mab Pwa-14, 2) immune mouse serum, 3) infected human serum against P. westermani extract and 4) the protein band of P. westermani extract on a nitrocellulose paper, stained with Amido-Black B. Antigenic bands of 28 kDa, 42.5kDa, 89 kDa and 120.5 kDa reacted on Pwa-14 were identified.


	Fig. 3 Localization of Mab by indirect fluorescent antibody technique. Frozen sections of P. westermani adult worm was reacted with: (A) Pwa-14 Mab showing positive reactions at the vitelline follicles and (B) immune mouse serum as a positive control (×100). (V: vitelline follicle, T: tegument).


	Fig. 4 Distribution of absorbance of human sera in paragonimiasis (Pw), clonorchiasis (Cs), cysticercosis (Cy), sparganosis (Sp) cases and normal controls (NHS) against P. westermani extract by micro-ELISA.


	Fig. 5 Inhibition rate (%) of Mab by human sera in the ELISA-inhibition test using P. westermani specific Mab Pwa-14. The cut-off value is set at 15%.

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