Description

The Diagnostic Automation, Inc Mumps IgM ELISA kit is intended for the qualitative detection of IgM antibody in human serum to Mumps for the determination of immunological experience. The performance characteristics of this kit have not been established. High complexity test.

The mumps virus is a member of the paramyxovirus group and the etiological agent of mumps in man. Mumps is a generalized illness usually accompanied by parotid (salivary gland) swelling and mild symptoms. It is also one of the most common causes of aseptic meningitis, encephalitis, and inflammation of the testes (orchitis), pancreas, and ovaries.

Parotitis as a presenting symptom in mumps infections is usually sufficiently diagnostic to preclude serological confirmation. However, a third of mumps infections are subclinical or unrecognized (1) and may require viral isolation and/or some other serological procedure to confirm or rule out mumps infection. An example of this is presenting orchitis or meningoencephalitis, the two most common complications of mumps infection, without salivary gland involvement. Virus isolation is time consuming and cumbersome and is usually an impractical procedure for the typical clinical laboratory. Current methods for serodiagnosis of mumps infections are in-vitro serum neutralization, hemagglutination-inhibition (HAI), indirect immunofluorescence, and complement fixation (CF) tests. Of these methods, neutralization is reportedly the most specific. However, the neutralization test requires 4-5 days to complete the test. HAI and CF are reportedly less sensitive than the neutralization test. These methods lack specificity, which limits their usefulness in determining immune status. The HAI test also requires pretreatment of test sera to remove nonspecific hemagglutination inhibitors from some sera.

Infection with mumps virus, whether symptomatic or subclinical, is generally thought to offer lifelong immunity. Anti-Mumps virus IgM appear 2-3 days after the occurrence of the first clinical symptoms (these remain 2-3 months), followed by the production of Mumps IgG antibodies which persist lifelong following vaccination with live virus there is a seroconversion in 90% of cases, however, the titre is somewhat lower than in normal infections.

As first described by Engvall and Perlmann and Van Weeman (5), Enzyme Immunoassays can be both specific and sensitive for the detection and measurement of serum proteins. The sensitivity, specificity, and reproducibility of enzyme-linked immunoassays can be comparable to other serological tests for antibody, such as immunofluorescence, complement fixation,
hemagglutination and neutralization.

Enzyme-Linked Immunosorbent Assays (ELISA) rely on the ability of biological materials, (i.e., antigens) to adsorb to plastic surfaces such as polystyrene (solid phase). When antigens bound to the solid phase are brought into contact with a patient\\\'s serum, antigen specific antibody, if present, will bind to the antigen on the solid phase forming antigen- antibody complexes. Excess antibody is removed by washing. This is followed by the addition of goat anti-human IgM globulin conjugated with horseradish peroxidase which then binds to the antibody-antigen complexes. The excess conjugate is removed by washing, followed by the addition of chromogen/substrate, tetramethylbenzidine (TMB). If specific antibody to the antigen is present in the patient\\\'s serum, a blue color develops. When the enzymatic reaction is stopped with 1N H2SO4, the contents of the wells turn yellow. The yellow color, which is proportional to the concentration of antibody in the serum, can be read on a suitable spectrophotometer or ELISA microwell plate reader. The sensitivity, specificity, and reproducibility of ELISAs can be comparable to other serological tests for antibody, such as immunofluorescence, complement fixation, hemagglutination and radioimmunoassays.

ELISA is as sensitive as the neutralization test and more sensitive than CF and HAI which makes it a reliable test for determination of immune status. The DAI Mumps IgM ELISA kit provides all the necessary reagents for the rapid determination and quantitation of IgM antibody to mumps virus in human sera.