| Home | E-Submission | Sitemap | Contact us
top_img
Korean J Parasitol > Volume 27(1):1989 > Article

Original Article
Korean J Parasitol. 1989 Mar;27(1):9-14. English.
Published online Mar 20, 1994.  http://dx.doi.org/10.3347/kjp.1989.27.1.9
Copyright © 1989 by The Korean Society for Parasitology
Demonstration of species-specific and cross reactive components of Paragonimus westermani crude worm antigen by EITB
K H Joo,H Ahn,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

Enzyme-linked immunoelectrotransfer blot (EITB) using crude worm antigen of adult Paragonimus westermani was performed for human patients sera to identify the species-specific components. Crude antigen was obtained by homogenizing and centrifuging 24-week old adult worms at 10,000 rpm for 60 minutes in phosphate buffered saline (PBS, pH 7.2) containing phenyl methyl sulfonyl fluoride (PMSF). Gradient sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) was performed and blotted electrophoretically onto a sheet of nitrocellulose paper. The sheet was cut into strips and exposed to sera diluted 1: 200 with PBS. SDS-PAGE showed 26 protein bands ranging 229 to 10 kDa. Of them 229, 91, 60, 50, 35-31, 27, 25, 21, 17, 11 and 10 kDa components showed positive reaction with serum antibody of patients with P. westermani. Sera of patients infected with Clonorchis sinensis reacted with 35-31, 19, and 11 kDa bands. Human sera from cysticercosis and diphyllobothriasis cases showed non-specific cross reactions with 229, 35-31, 27, 25 and 17 kDa bands. Protein bands of 91, 60, 21 and 10 kDa showed strong positive reaction without cross reactions with sera from other helminthic infections.

Figures


Fig. 1
Silver-stained SDS-PAGE of P. westermani. The two outside lanes(HMW, LMW) contained molecular weight protein standards given in kilodaltons.


Fig. 2
Location of antibody binding bands of soluble extract o adult P. westermani. Serum from pationts infected with following parasites were investigated. 1~5: paragonimiasis, 6~10: clonorchiasis, 11~13: diphyllobothriasis, 14~16: taeniasis, 17~19: cysticercosis, 20~24: non-infected sera, 25: PBS.

References
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.
 
3. Choi SK, et al. Korea Univ Med J 1984;21(1):33–40.
4. Choi WY, Lee OR. [Agar-Gel Precipitin Reactions In Experimental Paragonimiasis]. Korean J Parasitol 1981;19(2):101–108.
 
5. Choi WY, Lee WK, Lee OR. [Indirect Fluorescent Antibody Test For Diagnosis Of Paragonimiasis]. Korean J Parasitol 1975;13(2):152–158.
 
6. 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.
 
7. 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.
 
8. 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.
 
9. Itoh M, et al. Jpn J Parasitol 1988;37(5):347–352.
11. Jin SW, et al. Korea Univ Med J 1983;20(1):191–199.
12. Kim DC, et al. Rep NIH Korea 1982;19:109–114.
13. 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.
 
14. Kim SI, Ko EK, Kang SY, Cho SY. [Antigenicity of the soluble egg antigen of Paragonimus westermani]. Korean J Parasitol 1986;24(1):49–54.
 
15. Lee OR. J Soonchunhyang Univ 1986;9(2):239–246.
16. 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.
 
17. Merril CR, et al. Science 1981;221:1437–1438.
 
18. Neville DM Jr. Molecular weight determination of protein-dodecyl sulfate complexes by gel electrophoresis in a discontinuous buffer system. J Biol Chem 25;246(20):6328–6334.
19. 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.
 
20. Slemenda SB, Maddison SE, Jong EC, Moore DD. Diagnosis of paragonimiasis by immunoblot. Am J Trop Med Hyg 1988;39(5):469–471.
 
21. Soh CT, et al. Yonsei Rep Trop Med 1985;16(1):1–10.
22. 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.
  
23. Yogore MG, et al. Am J Trop Med Hyg 1965;14:586–591.
24. Yokogawa M, et al. Jpn J Parasitol 1962;11(2):117–122.
Editorial Office
c/o Department of Medical Environmental Biology
Chung-AngUniversity College of Medicine, Dongjak-gu, Seoul 06974, Korea
Tel: +82-2-820-5683   Fax: +82-2-826-1123   E-mail: kjp.editor@gmail.com
About |  Browse Articles |  Current Issue |  For Authors and Reviewers
Copyright © 2018 by The Korean Society for Parasitology and Tropical Medicine. All rights reserved.     powerd by m2community