Studies on the Applicability of Various Antigen Preparations in Immunoblot Diagnosis of Paragonimiasis

폐흡충증 진단에 있어서 각종 항원의 적용성

A systematic study was conducted to identify and isolate a serologically pertinent antigen with high specific activity and low cross reactivity from P.westermani antigen. Differential centrifugation of the homogenate yield three particulate and one soluble fractions ; the $480{\times}G$ pellet(Pw1), the $7650{\times}G$ pellet(Pw2) treated with n-butanol(Pw3), and $100000{\times}G$ supernatnat(Pw4). Comparison of antigenicity of these antigens, based upon differential centrifugation, to that of saline extract of P. westermani 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 using Pwl. 2) Silver stained and SDS-PAGE of each antigen showed 34 and 13Kd 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, 10Kd band was observed as specific band. In conclusion, Pw3($100000{\times}G$ pellet urea soluble, treated with n-butanol) and Pw4($100000{\times}G$ supernatant) were usable for ELISA and immunoblot technique.