Monoclonal antibodies for rapid detection of viral antigens in shell vials

Abstract

Diagnosis of Cytomegalovirus Infections and Development of Monocional Antibodies Cytomegalovirus (CMV) is a common infection that can cause serious disease in infants who have acquired the virus in utero, in patients immunosuppressed for organ transplantation, and in other individuals who receive chemotherapy or high levels of corticosteroids for treatment of neoplastic and other medical processes. Only 4 years ago, the standard method for detecting CMV infection in the laboratory was the inoculation of diploid fibroblast cell cultures in tubes with subsequent identification of typical cytopathic effects that required an average time period of 7 8 days for recognition (3). Immunologic methods for rapid detection of CMV in clinical specimens waned for lack of high quality homotypic antisera that could produce sensitive and specific results and, importantly, be commercially available. Subsequently, two monoclonal antibodies were prepared against an early and a late antigen of the ADI69 strain of CMV by Shuster et al. at the Mayo Clinic. These monoclonal antibodies provided the key to the development of a rapid test for detection of the virus (10). Since 1973, we have used shell vials to diagnose Chlamydia trachomatis infections. This bacterium is .only 3 4 times larger than the CMV virion; nevertheless, centrifugation of the specimen inoculum at low speed (700 x g) has been shown to enhance the rate and ultimate infectivity of this organism and viruses in susceptible cell cultures. Because we wanted to be able to detect CMV as rapidly as possible after inoculation of cell cultures in the laboratory, we used the monoclonal antibody prepared to an early antigen (72,000 d) to assay for the viral components synthesized after 16 hr incubation by the indirect immunofluorescence test (monoclonal antibody available from DuPont Specialty Diagnostics, Wilmington, DE).