Detection of tetanus antitoxin using Eu(3+)-labeled anti-human immunoglobulin G monoclonal antibodies in a time-resolved fluorescence immunoassay.
Tetanus antitoxin in human sera was detected with solid-phase immunoassays in microtitration modules coated with tetanus toxoid by using Eu(3+)-labeled anti-human monoclonal antibodies on the basis of an exactly calibrated antibody standard. The use of a time-resolved fluorescence immunoassay (TR-FIA) significantly improved the quantitative detection of tetanus antitoxin over that of the enzyme-linked immunosorbent assay (ELISA) technique because of its high sensitivity and its wide measurement range, detecting antibody levels between 0.001 and 12.5 IU/ml with a single serum dilution of 1:100. For the same purpose, two different serum dilutions (1:100 and 1:1,000) were needed in the ELISA technique. TR-FIA is reproducible and can be performed in 3.5 h. A study of 2,630 serum samples was undertaken to examine the age-dependent distribution of titer levels, indicating the decline of sufficient protection in patients older than 60 years. The wide measurement range of TR-FIA enabled fast examination of large numbers of serum samples without the need for repetition, with further sample dilution, as was often necessary in the ELISA procedure.
Schröder
J P
JP
Department of Immunology, Ernst-Rodenwaldt-Institute, Koblenz, Federal Republic of Germany.
Kuhlmann
W D
WD
D000293:Adolescent; D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D000911:Antibodies, Monoclonal; D002648:Child; D002675:Child, Preschool; D004797:Enzyme-Linked Immunosorbent Assay; D005063:Europium; D005069:Evaluation Studies as Topic; D015200:Fluoroimmunoassay / Q000379:methods*; D006801:Humans; D007074:Immunoglobulin G; D007223:Infant; D008297:Male; D008875:Middle Aged; D012680:Sensitivity and Specificity; D013743:Tetanus Antitoxin / Q000097:blood*
D000911:Antibodies, Monoclonal; D007074:Immunoglobulin G; D013743:Tetanus Antitoxin; D005063:Europium