Preparation and Application of Recombinant Paragonimus Westermani (Pw) Adult Antigen

Objective: To purify recombinant Paragonimus Westermani(Pw) adult antigen expressed in bactieria Pw-1/pRSET B/BL 21. After renature, the purified products were applied to detect the sera collected from paragonimiasis patients, other parasitosis and normal persons with ELISA and the specificity, sensitivity and application prospect of ELISA using Pw were evaluated. Methods: After primary purification of the recombinent protein by dialyse, the primary purified protein was further purified by Fast Protein Liquid Chromatography(FPLC), then the effects of the two steps of purification methods were compared. The primary purified products by Oxidation/reduction GSH renatured were to abtain the recombinant protein antigen. Finally, the sera collected from patients of paragonimiasis, schistosomiasis, clonorchiasis sinensis, cysticerciasis, echinococcosis and normal persons were detected for corresponding antibodies against the recombinant protein antigen and the Pw adult crude antigen respectively by ELISA. Results: FPLC, the sensitivity of the recombinant antigen tested by paragonimiasis patients'sera was 91.07%, and there was not any cross reaction with the sera collected from other parasitosis and normal persons. In contrast, the sensitivity of crude antigen was 100%, while the percentages of cross reaction were as follows: schistosomiasis 11.43%, clonorchiasis sinensis 4.84%, cysticerciasis 3.22%, echinococcosis 5.88%, normal person 2.5%. Conclusion: The primary purified antigen applied in the immunodiagnosis has higher specificity.