INTENDED USE:
The Mouse Anti-Tetanus Toxoid IgG ELISA Kit detects and quantifies tetanus toxoid/toxin-specific IgG in mouse serum or plasma of vaccinated or immunized animals. This immunoassay is suitable for:
-Determining immune status relative to non-immune controls;
-Assessing efficacy of vaccines, including dosage, adjuvantcy, route of immunization and timing;
-Qualifying and/or standardizing vaccine batches and protocols.
GENERAL INFORMATION:
Tetanus, a neurotoxic disease resulting in severe muscular spasm, is caused by Clostridium tetani bacteria. Tetanus toxin, a protein exotoxin produced only by C. tetani, is central to tetanus pathogenesis. Vaccination with the inactivated toxin (toxoid) elicits high levels of protection from the disease. The toxoid also acts as an adjuvant to boost immune responses to other antigens when combined in a vaccine, either mixed or covalently conjugated. Vaccines for tetanus are available in combination with other vaccines, e.g., pertussis, diphtheria, H. influenza b, hepatitis & polio. Monitoring the efficacy of vaccines by determining the antitetanus levels in the host, including for clinical trials using new formulation of vaccines, is often required. The ADI Anti- Tetanus Toxoid ELISAs will quantify antibodies produced by vaccines as well as from infection with the toxin-producing organisms.
PRINCIPLE OF THE TEST:
The Mouse Anti-Tetanus Toxoid IgG ELISA kit is based on the binding of mouse anti-tetanus toxoid IgG in samples to tetanus toxoid immobilized on the microwells, and anti-tetanus toxoid antibody is detected by anti-mouse IgG-specific antibody conjugated to HRP (horseradish peroxidase) enzyme. After a washing step, chromogenic substrate (TMB) is added and color is developed by the enzymatic reaction of HRP on the substrate, which is directly proportional to the amount of anti-tetanus toxoid IgG present in the sample. Stopping Solution is added to terminate the reaction, and absorbance at 450nm is then measured using an ELISA microwell reader. The activity of mouse IgG antibody in samples is calculated relative to antitetanus toxoid calibrators.
PRODUCT SPECIFICATIONS:
Specificity
Purified tetanus toxoid is used to coat the microwells; thus the
assay is specific for antibodies directed to tetanus toxoid or toxin.
The anti-Mouse IgG HRP conjugate reacts with mouse IgG
antibodies that bind to toxoid on the plate; IgA, IgM and IgE
antibody would not be measured above background signals.
Assay Sensitivity
The tetanus toxoid coating level, HRP conjugate concentration
and Low NSB Sample Diluent are optimized to differentiate antitetanus toxoid IgG from background (non-antibody) signal with
mouse serum samples diluted 1:100.
Calibrator Values
The calibrators are dilutions of antibody reactive to tetanus
toxoid. Values are assigned as arbitrary anti-tetanus toxoid
activity units.