Adenovirus IgM ELISA

Enzyme immunoassay for the qualitative determination of IgM class antibodies against Adenovirus in human serum or plasma (citrate). Adenoviruses are double-stranded DNA viruses of about 70-90 nm lacking an envelope. The capsid contains 252 capsomeres and shows icosahedral symmetry. The capsomeres consist of hexons, pentons and fiberprotein trimers which are responsible for the induction of group- and type-specific antibodies. For the first time adenoviruses were isolated in 1953 from tonsils and adenoid tissue by Rowe. More than 80 adenoviruses are known at present. 47 out of them are pathogenic for men. They cause several diseases of different organic systems, mainly eyes, pharynx, respiratory and gastrointestinal system. Adenovirus infections are common and frequent. Most infections appear during childhood. They pass of latently so that the virus can still be detected in tonsils after two years. It is excrete via saliva and faeces. Gate to body are mouth, nasal pharynx and conjunctiva of the eye. Most infections pass off without symptoms. Around 5 % of all coughs and sneezes of children are caused by adenoviruses. Epidemics may occur in populations crowded together, for example acute respiratory disease in military groups, pharyngoconjunctival fever in swimming pools, and epidemic keratoconjunctivitis in medical facilities. Infection of hospitals and swimming pools gratify special demands on hygienics. The qualitative immunoenzymatic determination of IgM-class antibodies against Adenovirus is based on the ELISA (Enzyme-linked Immunosorbent Assay) technique. Microtiter strip wells are precoated with Adenovirus antigens to bind corresponding antibodies of the specimen. After washing the wells to remove all unbound sample material horseradish peroxidase (HRP) labelled anti-human IgM conjugate is added. This conjugate binds to the captured Adenovirus-specific antibodies. The immune complex formed by the bound conjugate is visualized by adding Tetramethylbenzidine (TMB) substrate which gives a blue reaction product. The intensity of this product is proportional to the amount of Adenovirus specific IgM antibodies in the specimen. Sulphuric acid is added to stop the reaction. This produces a yellow endpoint colour. Absorbance at 450 nm is read using an ELISA microwell plate reader.

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