Bordetella pertussis IgM ELISA
The IMMUNOLAB Bordetella pertussis IgM Antibody ELISA Test Kit has been designed for the the detection and the quantitative determination of specific IgM antibodies against Bordetella pertussis in serum and plasma. Further applications in other body fluids are possible and can be requested from the Technical Service of IMMUNOLAB .
Whooping cough is a disease of the respiratory tracts which is caused by Bordetella pertussis bacteria. It is transmitted by airborne infection. The gramnegative Coccobacillus produces a series of biologically active molecules. The different compounds appear either during the pathogenesis or during the process of immunization against pertussis and show different effects. A characterisation has been made for the pertussis toxin (pt), the filamentery haemagglutinine (fha) and different lipopolysaccharides (lps).
Pertussis shows a high rate of transmission (rates of infection of over 90 % have been found for non-vaccinated household members) and can cause severe diseases, especially for very young children. From 10749 patients under one year between 1980 and 1989 69 % were brought into hospital, 22 % suffered from pneumonia, 0.9 % showed an Encephalopathy and 0.6 % died. For older children and adults (including already vaccinated persons) the infection may be observed by an unspecified bronchitis or inflammation of the upper respiratory tracts. Even asymptomatic cases are quite common.
The serological response following pertussis disease or immunization with pertussis vaccine has been measured with agglutination assays, precipitins, complement fixation and enzyme-linked immunosorbent assay (ELISA).
Enzyme-linked immunosorbent assays, in which Bordetella antigen (containing toxin, FHA and LPS and standardized in U/ml) is bound to a solid phase support, are sensitive, easy to perform and can be used both to determine seropositivity with a single serum and to indicate recent Bordetella infection by determination of IgM and IgA.
The IMMUNOLAB Bordetella pertussis IgM antibody test kit is based on the principle of the enzyme immunoassay (EIA). Bordetella 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 IgM antibodies of the serum and the immobilized Bordetella 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-IgM 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 IgM antibodies is directly proportional to the intensity of the color.
- Chodorowska, M. et al. ELISA test used for serologic diagnosis of Pertussis. Med. Dosw. Microbiol., 48: 15 (1996).
- Finger, H. et al. Serological diagnosis of whooping cough. Dev. Biol. Stand., 610: 331 (1985).
- Granström, G. et al. Specific Immunoglobulin A to bordetella pertussis antigen. J. Clin. Microbiol. 26: 869 (1988). 4
- Kuno-Sakai, H. et al.: A simple and sensitive ELISA of antibodies to Pertussis antigens. Vacine 10: 350 (1992).
- Nagel, J. et al.: Improved serodiagnosis of whooping cough caused by Bordetella pertussis. Dev. Biol. Stand. 610: 325 (1985).
- Reizenstein, E. et al.: Comparison of five calculation modes for antibody ELISA against Pertussis. J. Immunol. Methods 183: 279 (1995).
- Sato, Y. et al.: An improved ELISA system for the measurement of IgG antibodies against pertussis. Dev. Biol. Stand. 73: 167 (1991).
- Steketee, R. W. et al.: A comparison of laboratory and clinical methods for diagnosing pertussis. J. Infect. Dis. 157: 441 (1988).