A sensitive Enzyme-Linked Immunosorbent Assay(ELISA) for Vibrio vulnificus cytolysin was established.
Cytolytic activity in V. vulnificus culture supernatant reached to the maximum at about 4 hours and decreased rapidly afterwards. V. vulnificus cytol...
A sensitive Enzyme-Linked Immunosorbent Assay(ELISA) for Vibrio vulnificus cytolysin was established.
Cytolytic activity in V. vulnificus culture supernatant reached to the maximum at about 4 hours and decreased rapidly afterwards. V. vulnificus cytolysin was completely purified from culture supernatants by ammonium sulfate precipitation, DEAE-cellulose chromatography and phenyl-Sepharose chromatography. The polyclonal antibodies against the purified cytolysin were produced from a chicken and a rabbit. Resulting antibodies showed a high antigen-specificity for cytolysin, visualizing a band of 51 kDa when the V. vulnificus culture supernatants were on immunoblottings. Anti-cytolysin polyclonal rabbit and chicken antibodies were used as a capture or a detector antibody in sandwich ELISA after further purification with cytlysin-AFFi-prep immunoaffinity chromatography. The limit of detection was 10 ng of V. vulnificus cytolysin, with saturation achieved at 50 ng.