Quantitation of anti‐tetanus and anti‐diphtheria antibodies by enzymoimmunoassay: Methodology and applications

Abstract

We have developed enzymoimmunoassays (EIA) for the quantitation of antibodies (Ab) to tetanus and diphtheria toxoids (TT, DT) using Immulon I plates coated with the appropriate toxoid. A preparation of human tetanus immunoglobulin with a known concentration of anti-TT Ab was used as calibrator of the anti-TT antibody assay. The assay of anti-DT Ab is calibrated with a pool of human sera whose anti-DT Ab concentration was determined by quantitative immunoelectrophoresis, using a horse anti-DT with known Ab concentration as calibrator. A peroxidase-conjugated anti-human IgG was used in both assays. ABTS was used as substrate, and the reaction was stopped after 1 min incubation with citric acid and the OD measured at 414 nm on a Vmax reader. The assays have been applied to a variety of clinical situations. In patients suspected of having tetanus, the quantitation of antibodies has been helpful in establishing a diagnosis. In patients with a history of hypersensitivity to tetanus toxoid, verification of the levels of anti-TT antibody may prevent unnecessary and potentially harmful immunizations. The assays have also been used for the diagnostic evaluation of the humoral immune response to TT and DT, both in pediatric patients and in immunosuppressed patients. Several non-responders have been detected, and we have recently used the assay to monitor the effects of fish oil administration on the humoral immune response.(ABSTRACT TRUNCATED AT 250 WORDS)

Cellular and humoral immune responses to a tetanus toxoid booster in perinatally HIV-1-infected children and adolescents receiving highly active antiretroviral therapy (HAART)

“…Anti-tetanus toxoid immunoglobulin G (IgG) was measured using the Enzyme Immunoassay (EIA) Kit (BINDAZYME, The Binding Site Ltd., Birmingham, UK). A protective TAb was defined as ≥0.15 IU/ml, as described by previous studies for this assay [5,11,15,19,20].…”

Safety and immunogenicity of concomitant versus nonconcomitant administration of hepatitis B, tetanus-diphtheria, and measles-mumps-rubella vaccines in healthy eleven- to twelve-year-olds

“…Diphtheria antibody was tested by serum neutralization of diphtheria toxin in VERO cultures (cell cultures using green monkey kidney cells) [2,4]. Tetanus, measles, mumps, and rubella antibodies were assayed by enzyme immunoassay EIA [5][6][7]. Hepatitis B surface antibodies were measured quantitatively by a commercially available assay (Abbott Laboratories, Abbott Park, Illinois).…”

Approaches to Detecting Immunotoxic Effects of Environmental Contaminants in Humans

“…The anamnestic immune response can be evaluated by determining serum antibody levels to antigens that humans are commonly vaccinated with, such as tetanus toxoid (TT) and diphtheria (54). The levels of protective antibody specific for these antigens in the population vary with age.…”