Microtiter enzyme-linked immunosorbent assay for immunoglobulin G cholera antitoxin in humans: method and correlation with rabbit skin vascular permeability factor technique

Abstract

A microtiter enzyme-linked immunosorbent assay (ELISA) to measure immunoglobulin G cholera antitoxin in human serum has been developed. The ELISA employs commercially available reagents, including cholera enterotoxin and goat anti-human immunoglobulin G. It is specific, sensitive, and reproducible and requires as little as 5 microliter of serum. ELISA, moreover, permits quantitative determination of cholera antitoxin at a single serum dilution of 1:200. A total of 162 pre- and post-challenge sera from 49 volunteers who ingested Vibrio cholerae classical biotype, and 165 sera from 43 volunteers who ingested V. cholerae El Tor biotype, were tested for cholera antitoxin by ELISA and by the rabbit skin vascular permeability factor assay. The correlation between the two assays was statistically significant (P less than 0.001). ELISA for immunoglobulin G cholera antitoxin thus provides a valuable in vitro correlate of in vivo toxin-neutralizing capacity. Microtiter ELISA permits duplicate evaluation of at least 14 sera per 96-well plate including blanks and controls, is readily adapted to use in field studies, and therefore is particularly well suited to seroepidemiological surveys.