Evaluation of various microtiter plates with measles and gamma globulin antigens in enzyme-linked immunosorbent assays

 

Evaluation of various microtiter plates in enzyme-linked immunosorbent assays for antibodies to measles virus and to human immunoglobulin.

 

Measles virus is a negative-stranded enveloped RNA virus of the genus Morbillivirus in the family Paramyxoviridae that infects only humans and subhuman primates (Kreis et al., 1997; Fields et al. 1996). The virus is highly pleomorphic with a diameter varying between 150 and 300 nm. The envelope spikes of measles virus contain two viral glycoproteins: the hemagglutinin (H) and fusion (F) protein. The H is a type II transmembrane glycoprotein responsible for the binding of measles virus to cells, and the F protein is responsible for the uptake of virus into the cell. Antibodies to H and F are both required to neutralise the virus (Rollag and Haukenes, 1989).

Measles is normally a mild, highly contagious disease contracted by children (Rollag and Haukenes, 1989), and the infection with measles virus, as well as immunisation, induces a life-long protective immune reaction (Muller et al., 1993). In Third World countries, however, due to malnourishment, mortality can still be as high as 80%. Nevertheless, the world-wide morbidity and mortality of measles has fallen steadily since vaccination commenced in the early 1960’s (Dimmock and Primrose, 1994). Complete eradication of the disease, however, relies on simple, inexpensive, rapid, and yet very accurate measles diagnosis. Control of measles virus still causes serious problem, especially in developing countries, and it is important to design tests that can be used under field conditions. (Bouche et al., 1998a).

Enzyme-linked immunosorbent assay (ELISA) is a rapid, simple, reproducible, and rather inexpensive method that can be used for routine diagnosis of measles infection and immunisation (Kahane et al., 1979). The antigen used to measure the quantity of anti-MV antibody present in a sample can be either whole measles virus or recombinant measles H protein, or a peptide from the MV-H protein. As the H protein is the main target for neutralising and protective antibodies, H-ELISA tends to be more specific, more accurate, and more cost-efficient than whole-virus-based ELISA. Additionally, unlike whole MV-based antigens, H-protein could also be suitable for use in the development of a simple field test for the diagnosis of measles (Bouche et al., 1998b).

A major problem, both economically and environmentally, linked with the ELISA technique is that it requires the use of important quantities of polystyrene test-plates. Ways have already been found to regenerate and to reuse immunosorbent surfaces (Blanchard et al., 1990), but the use of biodegradable plates would be an additional way to reduce pollution.

The purpose of this project is to investigate the quality of different types of regular polystyrene and of biodegradable ELISA microtiter plates under various coating conditions.

 

Acknowledgements

I thank Dr. F. Schneider, Director of the Laboratoire National de Santé, Luxembourg, and Prof. Dr. C.P. Muller, Head of the Department of Immunology of the Laboratoire National de Santé, Luxembourg, for offering me the position as a summer-student at the Department of Immunology of the Laboratoire National de Santé, Luxembourg. I also thank F. Bouche and V. Bemtgen-Reddy from the Department of Immunology of the Laboratoire National de Santé, Luxembourg, for their support and for the excellent collaboration.

 

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