ABSTRACT
We screened echinococcosis-specific diagnostic antigen and verified its immunogenicity.The sequencing data of Echinococcus granulosus released by the National Human Genome Research Center was analyzed.The bioinformaties method was used to screen out Eg-07279 antigen gene Eg-07279,which was not expressed in Echinococcus granulosus and highly expressed in the original metacercariae.After expression,the recombinant protein rEg-07279 was purified by affinity chromatography.The specific IgG level was detected in the recombinant protein immunized mice and the immunogenicity was verified by Western blot.The selected antigen molecule Eg-07279 was cloned,expressed and purified to obtain the recombinant protein.The results of ELISA showed that the specific IgG level of Eg-07279 (2.559±0.125) was significantly higher than that of the blank group (0.319 0±0.01),the difference was statistically significant (P＜0.01);the Western blot results showed that the recombinant plasmids could be recognized by the secondary infection of the protoscolex and the serum of the recombinant protein immunized group,while the serum of the blank control group was not recognized by the serum.In summary,the Eg07279 antigen of E.granulosus was obtained and it was proved that the recombinant protein has good immunogenicity and is a good diagnostic antigen candidate molecule.
ABSTRACT
Leptospirosis, a zoonotic disease that is caused by many serovars which are more than 200 in the world, is an emerging worldwide disease. Accurate and rapid diagnostic tests for leptospirosis are a critical step to diagnose the disease. There are some commercial kits available for diagnosis of leptospirosis, but the obscurity of a species- or genus-specific antigen of pathogenic Leptospira interrogans causes the reduced sensitivity and specificity. In this study, the polysaccharide derived from lipopolysaccharide (LPS) of nonpathogenic Leptospira biflexa serovar patoc was prepared, and the antigenicity was confirmed by immunoblot and enzyme linked immunosorbent assay (ELISA). The performance of the rapid diagnostic test (RDT) kit using the polysaccharide as a diagnostic antigen was evaluated in Korea, Bulgaria and Argentina. The sensitivity was 93.9%, 100%, and 81.0% and the specificity was 97.9%, 100%, and 95.4% in Korea (which is a rare region occurring with 2 serovars mostly), Bulgaria (epidemic region with 3 serovars chiefly) and Argentina (endemic region with 19 serovars mainly) respectively. These results indicate that this RDT is applicable for global diagnosis of leptospirosis. This rapid and effective diagnosis will be helpful for diagnosis and manage of leptospirosis to use and the polysaccharide of Leptospira may be called as genus specific antigen for diagnosis.
Subject(s)
Female , Humans , Male , Antigens, Bacterial/immunology , Argentina , Bulgaria , Enzyme-Linked Immunosorbent Assay , Leptospira/isolation & purification , Leptospira interrogans/isolation & purification , Leptospirosis/diagnosis , Polysaccharides/immunology , Reagent Kits, Diagnostic/standards , Republic of Korea , Sensitivity and SpecificityABSTRACT
Leptospirosis, a zoonotic disease that is caused by many serovars which are more than 200 in the world, is an emerging worldwide disease. Accurate and rapid diagnostic tests for leptospirosis are a critical step to diagnose the disease. There are some commercial kits available for diagnosis of leptospirosis, but the obscurity of a species- or genus-specific antigen of pathogenic Leptospira interrogans causes the reduced sensitivity and specificity. In this study, the polysaccharide derived from lipopolysaccharide (LPS) of nonpathogenic Leptospira biflexa serovar patoc was prepared, and the antigenicity was confirmed by immunoblot and enzyme linked immunosorbent assay (ELISA). The performance of the rapid diagnostic test (RDT) kit using the polysaccharide as a diagnostic antigen was evaluated in Korea, Bulgaria and Argentina. The sensitivity was 93.9%, 100%, and 81.0% and the specificity was 97.9%, 100%, and 95.4% in Korea (which is a rare region occurring with 2 serovars mostly), Bulgaria (epidemic region with 3 serovars chiefly) and Argentina (endemic region with 19 serovars mainly) respectively. These results indicate that this RDT is applicable for global diagnosis of leptospirosis. This rapid and effective diagnosis will be helpful for diagnosis and manage of leptospirosis to use and the polysaccharide of Leptospira may be called as genus specific antigen for diagnosis.