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Korean J Parasitol > Volume 27(1):1989 > Article

Original Article
Korean J Parasitol. 1989 Mar;27(1):1-7. English.
Published online Mar 20, 1994.  http://dx.doi.org/10.3347/kjp.1989.27.1.1
Copyright © 1989 by The Korean Society for Parasitology
Analysis of antigenic specificities of Paragonimus westermani developmental stages using immunoblot technique
K H Joo,S C Hong,M S Chung and H J Rim
Department of Parasitology and Institute of Tropical Endemic Diseases, College of Medicine, Korea University, Seoul 110-702, Korea.
Abstract

Serodiagnosis of parasitic infections is widely used, since parasites or their eggs are not always detected by ordinary methods. The sensitive tests such as ELISA are highly dependent on the purity of antigens used. To solve this problem, many workers have tried to find species-specific components of antigens. The present study was performed to determine the antigenic profile of crude saline extracts of 3, 5, 8 and 12-week old P. westermani worms, which were collected from experimentally infected cats, based on SDS-PAGE and immunoblot technique. The results were as follows: 1. The SDS-PAGE showed at least 30 protein bands ranging from 229 kDa to 10 kDa molecular weight. The protein components of P. westermani changed chronologically during its developmental period. The 229 kDa band was recognized only in 12-week old worms (SEP12). 2. Analysis by ELISA showed a significant increase in antibody levels at 3 weeks in infected cats using crude saline extract antigens (SEP3, SEP5, SEP8, SEP12). 3. By EITB using SEP3 and SEP5, infected cats recognized major protein bands with molecular weight of 60, 35, 28, 25 or 21 kDa at 3-12 weeks of infection, and 3 additional antigens, 19, 13 and 10 kDa, were detected at 8-12 weeks of infections. 4. Using SEP8, 5 antigens, 91, 85, 31, 25 and 21 kDa, were consistently detected by all infected sera tested. In addition, 3 antigens of 19, 13 and 10 kDa were detected at 8-12 weeks of infection.(ABSTRACT TRUNCATED AT 250 WORDS)

Figures


Fig. 1
Silver-stained SDS-PAGE of saline extract antigen(SEP) of P. westermani.

(SEP3:antigen prepared from P. westermani worms collected from the cat at 3 weeks after infection, SEP5:at 5 weeks of infection, SEP8:at 8 weeks of infection, SEP12:at 12 weeks of infection, HMW: high molecular weight marker, LMW: low molecular weight marker, Kd, kDa: kilodaltons)



Fig. 2
Antigen-antibody binding patterns of sera from experimental cats against chronologically prepared P. westermani antigen. (Sera were collected at A: 3 weeks after infection, B: 5 weeks after infection, C: 8 weeks after infection, 12: weeks after infection)

Tables


Table 1
Chronological change of absorbance value of ELISA using crude antigen of P. westermani collected from cats at 3, 5, 8 and 12 weeks after infection


Table 2
Antigen-antibody reactions against major antigen bands in chronologically prepared P. westermani antigen (SDS-PAGE and EITB)

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