Diphtheria Toxoid IgG ELISA

The IMMUNOLAB Diphtheria Toxoid IgG Antibody ELISA Test Kit has been designed for the detection and the quantitative determination of specific IgG antibodies against Diphtheria Toxoid in serum and plasma. Further applications in other body fluids are possible and can be requested from the Technical Service of IMMUNOLAB.


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Product Catalog No: ILE-DIP01 Pack Size: 96 Wells

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Summary

Diphtheria is a bacterial infectious disease which appears predominantly during the childhood. The disease leads particularly to an inflammation of the pharynx, larynx and nasal mucosa. Additionally, bacterial toxins cause via long-distance effect damages of the heart, circulation and CNS. Only the toxigenic strains are pathogenic.

The etiologic agent is the Corynebacterium diphtheriae. These gram-positive bacteria prefer a microaerophil to anaerobe environment. Its pathogenicity is based on the secretion of an exotoxin that is circulating in the blood and effecting the heart muscle, kidneys and CNS. The Diphtheria toxoid will be produced by lysogenic strains.

Depending on the stage of disease, the three types ‘slight, middle and serious’ can be distinguished. The natural source of infection is the sick individual, whereas a carrier not absolutely shows symptoms. The infection is spread both through the aerial-droplet route and rarely by milk or smear infection. The appearance of Diphtheria shows a seasonal prevalence with the greatest incidence in winter. Especially non-vaccinated children will be infected. The incubation time is depending on the number of invasive germs.

The place of infection is the mucosa of the respiratory tract, where an acute local infection is developing. The secreted toxin leads to a superficial inflammation of the mucosa associated with the formation of a brown film (pseudo-membrane) upon it, consisting of bacteria, necrotic epithelial cells, fibrin, red and white cells. From this local inflammation, the toxin reaches other organs by using the blood and lymphatic circulation. Here it may cause severe damages. The grade of disease depends on the immunostate of the child. Usually, a limited Diphtheria arises, whereas in case of an immunosuppression, a severe Diphtheria is observed. As a result of this disease course, patients may die.

In most cases children will be vaccinated (e.g. DTP = Diphtheria-Tetanus-Pertussis) after the third month of life. The state of immunity can be monitored by determining the antitoxin IgG.

Test Principle

The IMMUNOLAB Diphtheria Toxoid IgG antibody test kit is based on the principle of the enzyme immunoassay (EIA). Diphtheria Toxoid antigen is bound on the surface of the microtiter strips. Diluted patient serum or ready-to-use standards are pipetted into the wells of the microtiter plate. A binding between the IgG antibodies of the serum and the immobilized Diphtheria Toxoid antigen takes place. After a one hour incubation at room temperature, the plate is rinsed with diluted wash solution, in order to remove unbound material. Then ready-to-use anti-human-IgG peroxidase conjugate is added and incubated for 30 minutes. After a further washing step, the substrate (TMB) solution is pipetted and incubated for 20 minutes, inducing the development of a blue dye in the wells. The color development is terminated by the addition of a stop solution, which changes the color from blue to yellow. The resulting dye is measured spectrophotometrically at the wavelength of 450 nm. The concentration of the IgG antibodies is directly proportional to the intensity of the color.

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    References
    • Borcic, B. et al. Immunity to diphtheria in the Croatian population in 1994. Lijec. Vjesn., 118: 227 (1996).
    • CDC. Diphtheria Epidemic – New Independent States of the Former Soviet Union. 1990-1994; MMWR 44: 177 (1995).
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    • Frank, J.W. et al. Diphtheria overimmunization in children. Can. Med. Assoc. J., 141: 1241 (1989).
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    • Kjeldsen, K. et al. Immunity against diphtheria and tetanus in the age group 30-70 years. Scand. J. Infect. Dis,. 20: 177 (1988).
    • Lau, R.C. Detection of diphtheria toxin antibodies in human sera by ELISA. J. Hyg., 96: 415 (1986).
    • Mueller, J.H. et al. Production of diphtheria toxin of high potency (100 Lf) on a reproducible medium. J. Immunol., 40: 21 (1941).
    • Krech, T., Naumann, P., Wittelsbürger, CH. et al. Dtsch. med. Wschr., 112: 541 (1987).
    • Melville-Smith, M. et al. Estimation of diphtheria antitoxin in human sera by ELISA. J. Med. Microbiol., 25: 279 (1988).
    • Rieger, J. et al. Diphtheria immunity in the German population; Gesundheitswesen, 56: 667 (1994).
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