Two-site monoclonal antibody quantitative ELISA for toxic shock syndrome toxin-1.

A two-site monoclonal antibody (MAB) quantitative enzyme-linked immunosorbent assay (ELISA) was developed that enables quantitation of toxic shock syndrome toxin-1 (TSST-1) down to 0.25 ng/ml and detection of TSST-1 to 0.06 ng/ml. Interference by Staphylococcus protein A was eliminated by incorporating normal rabbit serum into the test sample diluent. In the process of selecting an MAB pair for a two-site 'sandwich'-type ELISA, the MABs were screened for inhibition or common epitope binding. Some MABs that reacted with antigen that was adsorbed to a polystyrene well would not bind to antigen that was presented in a more natural configuration, as in the case of antigen immobilized by trapping antibody. Conversely, MABs that reacted with antigen that was immobilized by another antibody did not all function as trapping antibodies when adsorbed directly to a polystyrene surface. ELISAs that used polyclonal antibodies in the capture mode and MAB conjugate as the second antibody were generally more sensitive than were those that used polyclonal antibodies for both capture and indicator functions. MAB screening and selection schemes should be carefully designed to evaluate MABs in the mode in which they will be used in the final assay.

Antigenic and genetic variation in Legionella pneumophila serogroup 6.

Legionella pneumophila subsp. pneumophila serogroup 6 is second in importance only to L. pneumophila serogroup 1 as a cause of legionellosis. Monoclonal antibody (MAb) reactivity and multilocus enzyme electrophoretic analyses were used to subtype serogroup 6 isolates as a potential aid for epidemiologic and virulence studies. Forty-eight serogroup 6 isolates submitted to the Centers for Disease Control from 1980 to 1985 were examined by these methods. The isolates were divided into two groups based on differential reactivity with two MAbs. Thirty-two of the isolates were of a single electrophoretic type (ET) and were reactive with both MAbs. The remaining 16 isolates were distributed among 10 ETs and were reactive with one or both MAbs. The mean genetic diversity for serogroup 6, as determined from the degree of variability at 20 enzyme loci, was found to be essentially the same as that for L. pneumophila subsp. pneumophila as a whole. The ETs of serogroup 6 isolates were unique but closely related genetically to the ETs of L. pneumophila subsp. pneumophila serogroups 1 to 5, 7, and 8. The range of serogroup 6 subtypes distinguished by MAbs and enzyme electrophoresis suggests that the combination of these two methods can be useful as a typing system.