Attempts to increase the sensitivity of fluorescein-based fluorescence immunoassays by using multiple labelling have generally been unsuccessful because of concentration quenching. We have labelled antibodies to human immunoglobulin G with multiple fluorescein fluorophores attached by means of a disulphide linkage: this linkage can be rapidly and easily broken by treatment with dithiothreitol, allowing fluorescein to be released from the antibody and measured in free solution. Application of this technique to a fluorescence labelled immunosorbent assay for antibodies to the human immunodeficiency virus gave an approximately 20-fold increase in signal compared with an equivalent assay using fluorescein isothiocyanate.

The molecular weight distributions of resins produced by reaction of formaldehyde with phenols, melamine, or urea have been studied by gel permeation chromatography.

We describe a new immunoassay for the detection of antibodies to the human immunodeficiency virus. The method is based on a silver enhanced gold-labelled immunosorbent assay (SEGLISA). Test sera are incubated in microtitre wells on which antigens have been coated. If present in the test sera, antibodies to the human immunodeficiency virus bind to the solid-phase antigens. Bound antibodies are quantitated with anti-human immunoglobulin labelled with gold. Positive specimens produce a faint pink deposit which is better visualised by silver enhancement which gives an intense black colour. The intensity of the colour is proportional to the bound antibody concentration. All the reagents are stable and the silver enhancement takes place under ambient light conditions. The assay has many of the advantages of micro enzyme-linked immunosorbent assays but does not suffer from the drawbacks associated with the use of an enzyme label. It requires fewer manipulations and is quicker to carry out than an equivalent enzyme-linked test. As the silver layer is permanent dried wells may be stored for future reading or checking.

The development and assessment of a fluorescence-labelled immunosorbent assay for the detection of antibodies to the human immunodeficiency virus is described. Test serum is incubated in microtitre wells on which antigens have been coated. If present in the test serum, antibodies to the human immunodeficiency virus bind to the solid-phase antigens. In turn the antibodies are quantified with anti-human immunoglobulin labelled with fluorescein. Positive samples produce an intense fluorescence which is measured in a spectrofluorimeter. When used to test a panel consisting of normal serum and antibody-positive serum from infected patients the assay proved to be 100% specific and to have a sensitivity of 100%. The assay has many of the advantages of micro enzyme-linked immunosorbent assays, but does not suffer from the drawbacks associated with the use of an enzyme label. It requires fewer manipulations and is quicker to carry out than an equivalent enzyme-linked test.