Hari V, Baunoch D, Das P
Wayne State University, Detroit, MI 48202.
Biotechniques. 1990 Sep;9(3):342-50.
We have developed a sensitive method, sphere-linked immunodiagnostic assay, using specific antigens covalently bonded to microspheres for the detection of antibodies in serum. In this method, specific antigens, such as the capsid proteins of tobacco mosaic virus and tobacco etch virus, were independently, covalently bonded to plastic micropheres of 0.5 microns or 0.9 microns in diameter. The antigen-linked spheres were then exposed to normal serum or serum containing specific antibody, followed by treatment with gold-labeled secondary antibodies. The binding of the gold-labeled secondary antibodies to the specific primary antibodies on the spheres acted as an indication of the presence of the specific primary antibodies. The spheres were then examined and photographed by transmission electron microscopy. The number of gold particles bound to the spheres was counted manually using the photographs. The gold labeling was found to be specific and sensitive, enabling detection of antibodies present in highly diluted antisera. The efficiency and sensitivity of the technique for detection of antibodies were compared with those of the enzyme-linked immunosorbent assay and found to be highly sensitive. The technique was also used for testing for the presence of antibodies to herpes simplex virus as well as antibodies to Staphylococcus enterotoxin using microspheres coated with the respective antigens. We believe that this technique could be applied clinically when needed for detection of antibodies to other viruses, such as the Human Immunodeficiency Virus.
