Viral diagnosis by antigen detection techniques

Diagnosis of viral infections can be obtained in the early stages of a disease by detection of viral antigens directly in the clinical specimen. This has become an important tool for rapid virus diagnosis.

Antigens produced during virus infections can be detected either in cells collected from the site of infection by immunohistological investigation or in secretions and blood by solid phase immunoassays (IA). Viruses causing acute respiratory infections can be diagnosed in cells from the respiratory tract, viruses causing vesicular eruptions in epithelial cells from skin scrapings, rabies virus in nerve cells of the brain or epithelial cells from skin and cornea and cytomegalovirus (CMV) matrix antigen, pp65, can be detected in peripheral blood leukocytes (PBL) by immunofluorescence (IF) or immunoperoxidase techniques. The quality of specimens can be easily checked during the reading of results. Some IAs for antigen detection, such as detection of HBsAg and HIV p24 antigen in blood are standardized and sensitive. Others give less sensitive results because of the variation of quality of the clinical specimen. The latex agglutination tests are mainly used for rapid detection of virus or viral antigens in faeces: rota-and adenoviruses; the method may not be very sensitive but yields a result within a few minutes. Assays detecting viral nucleic acids are more sensitive than antigen detection tests because of a tremendous amplification of gene segments obtained by the polymerase chain reaction (PCR). So far such assays are time consuming and expensive and are mainly used in specific clinical situations.

After introduction of specific monoclonal antibodies (Mabs), the antigen detection techniques are increasingly used. the need for quality control, trained staff, and standardized reagents and methods for specimen collection and preparation is now being appreciated. IF for viral respiratory viruses is used for diagnosis and epidemiological studies all over the world. Likewise, IF is still the method most often used for rabies diagnosis. For CMV, the pp65 matrix antigen is shown to be a sensitive marker closely correlated with clinical symptoms. Its detection by the IF technique has proven to be superior to other techniques for prediction of CMV pneumonia in bone marrow transplant patients. IAs are currently used in fully automated systems for large scale diagnosis based on antigen detection in serum specimens. Increase of antibody specificity on the solid phase by use of Mabs directed against the most abundant viral antigen in the clinical specimen shortens the reaction time; this has been employed in most of the constantly appearing new rapid diagnosis kits based on the immunoassay principle.

Although, in virology, more sensitive results are obtained by the gene detection method, PCR, directly in clinical samples, viral antigen detection tests are, after the introduction of Mabs for diagnostic purposes, increasingly used because of their low demand on laboratory equipment, their rapid and early result and relatively low cost. Antigen detection is successfully used directly in clinical specimens for rapid diagnosis of many viral infections as well as for identification of tissue culture isolated viruses. With Mab-based IAs the reaction time is shortened and new rapid, almost 'instant test' kits are appearing on the market.