| Objective: To express the gene of Paragonimiasis westermani (Pw) pro- cathepsin L (Pwl) which have been converted into Escherichiacoli(BL21) and purify the expressed products . Using the purifiedproducts to test the sera of paragonimiasis patients, patients who haveother Parasitosis and normal persons with Enzyme-linked immunosorbentassays (ELI SAs) and evaluate its specificity, sensitivity andapplicationprospect. Methods: To induce the recombinantPw/pRESETB/E.coli BL21 expression by IPTG and analyse theexpressed recombinant protein with SDS-PAGE. To identify itsimmunological characteristics. The sera from the Pw adult antigenimmuned rabbits,nomal rabbits, Pw infected dog and normal dog,paragonimiasis patients and schistosomiasis patients were tested withWestern blot. Then,purified the expressed products in inclusion bodieswith step by step washing and dialysing. Renatured the purified productsby Oxidation / reduction GSH to abtain the recombinant protein antigen.Finally, the sera from the patients with paragonimiasis , schistosomiasis,clonorchiasis sinensis, cysticerciasis or echinococcosis respectively andnormal persons were detected for corresponding antibodies with therecombinant protein antigen and the Pw adult crude antigen , makingcomparison between these two antigens. Results: A specific proteinbelt was shown at about 32ku .The 32ku protein belt can only berecognised by the sera from the rabbits immunised with PwAg, the dogsinfected with Pw and the paragonimiasis patients. The ELISA resultsshowed that the recombinant protein antigen has no any cross reactionwith other parasitosis and normal persons. In contrast, the crudu antigen'scross reactions are as follows: schistosomiasis 11.43%, clonorchiasissinensis 4.84%, cysticerciasis 3.22%, echinococcosis5.88%, normalperson 6.67%. Conclusion: that the the specificity of the recombinant protein is very high, and can be applied to the immunodiagnosis ofparagonimisasis. |
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