A systematic study was conducted to identify and isolate a serologically pertinent antigen with high specific activity and low cross reactivity from Pwestermani antigen.
Differential centrifugation of the homogenate yield three particulate and one soluble fractions; the 48O X G pellet(Pwl ), the 7650 X G pellet(P w2), l00000 X G pellet treated with n-butanol(Pw3), and 100000XG supernatnat(Pw4). Comparison of antigenicity of these antigens. based upon differential centrifugation, to that of saline extract of P.westemwni worm(SEP) was performed by SDS-PAGE and immunoblot techniques.
The results obtained were as follows
1) The ratio of absorbance value of ELISA against paragonimiasis positive pool sera to that of negative sera was highest when using Pw3 as antigen, and that was lowest when using Pwl.
2) Silver stained and SDS-PAGE of each antigen showed 34 and 13 Kd band as common antigen band, but Pw2 didn’t show clear band.
3) By immunoblot. 55 and 34Kd bands using SEP and Pw4 showed strong positive reaction without cross reaction with sera from other helmenthic infections. Using Pw3, IOKd band was observed as specific band.
In conclusion. Pw3(100000XG pellet urea soluble, treated with n-butanol) and Pw4(100000 XG supernatant) were usable for ELISA and immunoblot technique.
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