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Parasit Host Dis > 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.

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)


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)


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)

1. Cho KM, et al. Yonsei Rep Trop Med 1976;7:26–37.
2. Cho SY, Hong ST, Rho YH, Choi SY, Han YC. Application of micro-ELISA in serodiagnosis of Human paragonimiasis. Korean J Parasitol 1981;19(2):151–156.
4. Choi WY, Yoo JE, Nam HW, Choi HR. [Purification of antigenic proteins of Paragonimus westermani and their applicability to experimental cat paragonimiasis]. Korean J Parasitol 1986;24(2):177–186.
5. Choi WY, Lee OR. [Agar-Gel Precipitin Reactions In Experimental Paragonimiasis]. Korean J Parasitol 1981;19(2):101–108.
6. Choi WY, Lee WK, Lee OR. [Indirect Fluorescent Antibody Test For Diagnosis Of Paragonimiasis]. Korean J Parasitol 1975;13(2):152–158.
7. Choi WY, Jin YK, Lee OR, Kim WG. [Analysis Of Protein Components At Varioue Stages Of Clonorchis Sinensis]. Korean J Parasitol 1981;19(1):8–17.
9. Hillyer GV, Serrano AE. The antigens of Paragonimus westermani, Schistosoma mansoni, and Fasciola hepatica adult worms. Evidence for the presence of cross-reactive antigens and for cross-protection to Schistosoma mansoni infection using antigens of Paragonimus westermani. Am J Trop Med Hyg 1983;32(2):350–358.
10. Huer B, Kim SI, Kang SY, Cho SY. Electrophoretic patterns of proteins from Paragonimus westermani in early developmental stages. Korean J Parasitol 1985;23(2):189–196.
11. Hunter GW 3rd, Ritchie LS, Pan C, Lin S, Sugiura S, Nagano K, Yokogawa M. Immunological studies, II. Intradermal tests and their application in the field for the detection of schistosomiasis japonica, paragonimiasis and clonorchiasis. Mil Med 1958;122(2):85–96.
13. Joo KH, Ahn H, Chung MS, Rim HJ. Demonstration of species-specific and cross reactive components of Paragonimus westermani crude worm antigen by EITB. Korean J Parasitol 1989;27(1):9–14.
15. Kim SI, Kang SY, Cho SY. [On The Applicability Of Partially Purified Antigenic Preparations Of Paragonimus Westermani]. Korean J Parasitol 1983;21(2):257–264.
18. Lee OR, Chang JK. [ELISA of paragonimiasis in cat by crude and purified antigens of Paragonimus westermani]. Korean J Parasitol 1986;24(2):187–193.
20. McLaren M, Draper CC, Roberts JM, Minter-Goedbloed E, Ligthart GS, Teesdale CH, Amin MA, Omer AH, Bartlett A, Voller A. Studies on the enzyme linked immunosorbent assay (ELISA) test for Schistosoma mansoni infections. Ann Trop Med Parasitol 1978;72(3):243–253.
21. Merril CR, Goldman D, Sedman SA, Ebert MH. Ultrasensitive stain for proteins in polyacrylamide gels shows regional variation in cerebrospinal fluid proteins. Science 27;211(4489):1437–1438.
22. Payares G, McLaren DJ, Evans WH, Smithers SR. Changes in the surface antigen profile of Schistosoma mansoni during maturation from cercaria to adult worm. Parasitology 1985;91(Pt 1):83–99.
23. Sadun EH, Buck AA, Walton BC. The diagnosis of paragonimiasis westermani using purified antigens in intradermal and complement fixation tests. Mil Med 1959;124(3):187–195.
24. Soh CT, et al. Yonsei Rep Trop Med 1985;16(1):1–10.
25. Towbin H, Staehelin T, Gordon J. Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc Natl Acad Sci U S A 1979;76(9):4350–4354.
26. Tsang VC, Peralta JM, Simons AR. Enzyme-linked immunoelectrotransfer blot techniques (EITB) for studying the specificities of antigens and antibodies separated by gel electrophoresis. Methods Enzymol 1983;92:377–391.
28. Yogore MG, et al. Am J Trop Med Hyg 1965;14:586–591.
29. Yokogawa M, et al. Jpn J Parasitol 1962;11(2):117–122.
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