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On the applicability of partially purified antigenic preparations of Paragonimus westermani
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Original Article
Korean J Parasitol. 1983 Dec;21(2):257-264. Korean.
Published online Mar 20, 1994.  http://dx.doi.org/10.3347/kjp.1983.21.2.257
Copyright © 1983 by The Korean Society for Parasitology
On the applicability of partially purified antigenic preparations of Paragonimus westermani
Suk-Il Kim,Shin-Yong Kang and Seung-Yull Cho
Department of Parasitology, College of Medicine, Chung-Ang University, Seoul 151, Korea.
Abstract

In order to obtain more specific antigenic preparation for the diagnosis of human paragonimasis, crude saline extract of whole worm (=PwWWE), secretory-excretory components (PwSEC) and secretion-excretion-free somatic extract (PwSM) of 12 week-old Paragonimus westermani were filtrated through Sephadex G-200 gel column.

The adult Paragonimus worms were obtained from experimentally infected dogs. A total of 11 antigenic solutions was lyophilized or diluted to adjust protein content of 1 mg/ml.

To evaluate the antigenicity of crude antigens and fractions, micro-ELISA was done with the sera from P. westermani infected cases, C. sinensis infected cases and non-infected control cases to detect Paragonimus specific IgG antibody.

The results were as follows:

1. When the PwWWE was filtrated through Sephadex G-200 gel, it was separated into three fractions; PwWWE Fr. 1, PwWWE Fr. 2 and PwWWE Fr. 3. The percentage of protein content was 28.0 percent, 21.6 percent and 50.4 percent respectively.

The PwSM was also separated into three fractions; PwSM Fr. 1, PwSM Fr. 2, PwSM Fr. 3 and their percentage of protein content was 41.3 percent, 38.6 percent and 20.1 percent.

However, the PwSEC showed different fractionation pattern; i.e. fraction 1 (=PwSEC Fr. 1) and 3 (PwSEC Fr. 3) without fraction 2. The percentage of protein content was 14.0 percent in PwSEC Fr. 1 and 86.0 percent in PwSEC Fr. 3.

2. When the antigenicity of each Paragonimus crude antigen and fractionated antigen was evaluated for specific IgG antibody by micro-ELISA in 10 human paragonimiasis sera, PwSEC Fr. 1 was the most potent antigen showing the mean absorbance 1.98. The PwWWE Fr. 1, PwSEC, PwWWE were next to that; their mean absorbance were 1.72, 1.38 and 0.83, respectively. The antigenicity of fractions 2 and 3 was much weaker in binding specific IgG antibody.

3. When the antigens were reacted in micro-ELISA with 10 human clonorchiasis sera, most antigens showed weak reactivity. Each fraction 1 of crude antigens reacted higher than other fractions or crude antigens; the mean absorbance was 0.17 in fraction 1, but in others the absorbances were about 0.06.

4. With non-infected control sera, the result of micro-ELISA revealed almost same pattern with those of the clonorchiasis sera.

From the above results, it became apparent that PwWWE Fr. 1, especially PwSEC Fr. 1 was the most potent antigen reacted with Paragonimus specifc IgG antibody.

Figures


Fig. 1
Sephadex G-200 gel filtration of saline extracts of adult P. westermani.

(a) P. westermani whole worm extract (PwWWE)

(b) P. westermani secretory-excretory components (PwSEC)

(c) P. westermani somatic extract (PwSM)


Tables


Table 1
Results of ELISA between the various antigenic preparations of P. westermani and the sera of 10 paragonimiasis cases


Table 2
Results of ELISA between the various antigenic preparations of P. westermani and the sera of 10 clonorchiasis cases


Table 3
Results of ELISA between the various antigenic preparations of P. westermani and the sera of 10 non-infected cases

References
1. Cho KM, et al. Yonsei Reports Trop Med 1976;7:26–39.
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 WY, Kimura K, Tsuji M. [Immunoelectrophoresis For Anthelmintics Evaluation Against Experimental Paragonimiasis]. Korean J Parasitol 1976;14(2):94–102.
 
4. Choi WY, Lee WK, Lee OR. [Indirect Fluorescent Antibody Test For Diagnosis Of Paragonimiasis]. Korean J Parasitol 1975;13(2):152–158.
 
5. 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.
 
6. Kim CH, et al. Chung-Ang J Med 1982;7:335–347.
7. Lowry OH, Rosebrough NJ, Farr AL, Randall RJ. Protein measurement with the Folin phenol reagent. J Biol Chem 1951;193(1):265–275.
 
8. Sadun EH, Walton BC, Buck AA, Lee BK. The use of purified antigens in the diagnosis of clonorchiasis sinensis by means of intradermal and complement fixation tests. J Parasitol 1959;45(2):129–134.
  
9. Sawada T, Takei K, Voneyama K. Studies On The Immunodiagnosis Of Paragonimiasis. I. The Precipitin Reaction With Crude And Fractionated Antigens. J Infect Dis 1964;114:311–314.
  
10. Tsuji M. Japanese J Parasit 1974;23:11–21.
11. Yogore MG, et al. Am J Trop Med Hyg 1965;14:586–591.
12. Yokogawa M, et al. Japanese J Parasit 1962;11:117–122.
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