| Home | E-Submission | Sitemap | Contact us |  
Korean J Parasitol > Volume 24(2):1986 > Article

Original Article
Korean J Parasitol. 1986 Dec;24(2):187-193. English.
Published online Mar 20, 1994.  http://dx.doi.org/10.3347/kjp.1986.24.2.187
Copyright © 1986 by The Korean Society for Parasitology
ELISA of paragonimiasis in cat by crude and purified antigens of Paragonimus westermani
Ok Ran Lee and Jae Kyung Chang
Department of Parasitology, Soonchunhyang University College of Medicine, Korea.
Department of Parasitology, College of Medicine, Yonsei University, Korea.

Enzyme-linked immunosorbent assay (ELISA) using crude and affinity-purified antigens of adult worms of Paragonimus westermani was performed for infected cat sera with different worm burden, from preinfection to 18th week after infection. Crude antigen was used with supernatant of homogenated worms by freezing-thawing method, and the supernate was centrifuged for l hour at 10,000 rpm at 4C. Affinity-purified antigen (antibody-bound antigen) was prepared from fractions (bound and unbound) of crude antigen by affinity chromatography on CNBr-activated sepharose 4B, and IgG as a ligand was prepared from paragonimiasis cat serum (6 months infected) obtained by ammonium sulfate (40-45 per cent saturated) precipitation method. By SDS-PAGE, crude antigen showed 22 polypeptide fractions while purified antigen showed 4 fractions: 36, 400, 34,700,27,600 and 11,500 in molecular weights. All cats were divided into five groups(G1-G5) by different worm burdens. The mean of recovered worms (±SD) and the number of cats in each group are as follows:G1, 2 worms(0) and 4 cats; G2, 4.75(±0.66) and eight; G3, 10.75(±1.92) and four; G4, 25.20(±3.43) and five; G5, 48(±12.63) and five cats. The results were summarized as follows: The antibody levels(OD value) increased by worm burden in G1 to G4 generally. However, individual antibody levels were not exactly related with worm burden in all groups, especially there was a wide difference in G4 and G5. These results suggested that the worm burden in G4 (about 20 - 30 worms) is enough to produce antibody maximum in cats of 2~3 kg weight. The antibody levels increased significantly (p<0.05) compared to control sera at the 3rd week in G1 and G2, at the 2nd week in G3, and at the 1st week in G4 and G5. Especially in the 4th week, OD value increased more in G1(p<0.001) and in G2 to G5(p<0.01). In the pattern of antibody levels by ELISA in each group, OD in G1 increased to the 18th week continuously, in G2 OD was maintained same after the 16th week, but in G3 it decresed after the 16th week, and it was maintained same in G4 and G5 after the 14th week. The antibody levels by ELISA with the affinity-purified antigen were higher than those with crude antigen in all groups generally. Especially, the difference of OD values between two antigens was larger from the 4th to the 10th week. In G1 and G2 OD with purified antigen was higher than that with crude one to the 18th week. It was also higher in G3 than that with crude antigen to the 16th week and OD of G4 and G5 were higher before the 14th week than that with crude antigen, however became lower at the 16th week. Consequently, the antibody level in ELISA with affinity-purified antigen was more sensitive at the early weeks after infection and in light infection groups than that with crude antigen.


Fig. 1
Affinity chromatography of crude worm extract of Paragonimus westermani using CNBr-activated Sepharose 4B.

Fig. 2
Electrophoretic analysis of crude and affinity-purified antigens of Paragonimus westermani

Fig. 3
Antibodies as detected by crude antigen of P. westermani; the specific IgG antibody titers in sequence from cats experimentally infected with worms of 2(G1), 4~6(G2), 9~14(G3), 18~28(G4), 34~69(G5) were measured suing ELISA.

Fig. 4
IgG antibody titer curves in ELISA of paragonimiasis westermani cat sera against crude and purified antigens of P. westermani


Table 1
Numbers of recovered worms in each group of experimental cat paragonimiasis

1. Choi WY, Lee OR. [Agar-Gel Precipitin Reactions In Experimental Paragonimiasis]. Korean J Parasitol 1981;19(2):101–108.
2. Han SH, Woo CK. [Immuno-Diffusion And Complement-Fixation Test In Paragonimiasis Of Cat]. Korean J Parasitol 1969;7(3):160–165.
3. Hillyer GV, Pelley RP, del Llano. Solubilization of antigens of Fasciola hepatica which react with antibodies to Schistosoma mansoni. J Parasitol 1979;65(1):55–60.
4. 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.
5. 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.
6. 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.
7. Hu XS, Feng RY, Hu AQ, Lin FQ, Yan HP, Tian GZ, Wang HX. Immunodiagnosis of paragonimiasis by counterimmunoelectrophoresis and agar gel diffusion. Chin Med J (Engl) 1980;93(8):557–561.
8. Imai JI. Tropical Med. (Nagasaki) 1979;21(2):45–55.
9. 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.
10. Laemmli UK. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 1970;227(5259):680–685.
11. Lee OR, Choi WY. [Comparison Of Agar-Gel Diffusion Tests, Counterimmunoelectrophoresis And Enzyme-Linked Immunosorbent Assay In The Sera Of Skin Test Positives For Paragonimiasis]. Korean J Parasitol 1983;21(2):270–280.
12. Lowry OH, Rosebrough NJ, Farr AL, Randall RJ. Protein measurement with the Folin phenol reagent. J Biol Chem 1951;193(1):265–275.
13. 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.
14. 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.
15. Sawada T, Takei K, Yoneyama K. Studies On The Immunodiagnosis Of Paragonimiasis. II. Intradermal Tests With Fractionated Antigens. J Infect Dis 1964;114:315–320.
16. Seed JR, et al. J Parasit 1966;52(2):358–362.
17. Tada I. Physiological and serological studies of Paragonimus miyazakii infection in rats. J Parasitol 1967;53(2):292–297.
18. Voller A, Bidwell DE, Bartlett A, Fleck DG, Perkins M, Oladehin B. A microplate enzyme-immunoassay for toxoplasma antibody. J Clin Pathol 1976;29(2):150–153.
19. Yokogawa M, et al. Jpn J Parasit 1983;32(4):317–322.
Editorial Office
Department of Molecular Parasitology, Samsung Medical Center, School of Medicine, Sungkyunkwan University,
2066 Seobu-ro, Jangan-gu, Suwon 16419, Gyeonggi-do, Korea.
Tel: +82-31-299-6251   FAX: +82-1-299-6269   E-mail: kjp.editor@gmail.com
About |  Browse Articles |  Current Issue |  For Authors and Reviewers
Copyright © 2022 by The Korean Society for Parasitology and Tropical Medicine.     Developed in M2PI