Detection of antibody to herpes simplex virus by enzyme linked immunosorbent assay technic using locally prepared antigen

Abstract

Serological methods are playing an increasingly important role in the diagnosis and epidemiological assessment of diseases. Simple and inexpensive methods for large-scale application are urgently needed. The enzyme linked immunoassay (ELISA) developed recently and reviewed elsewhere holds great promise in a wide variety of applications. In this study, an ELISA was developed for detecting antibody to herpes simplex virus (HSV). A crude extract of HSV-infected HeLa cells and non-infected cells were used to coat wells of polystyrene microtiter plates. Human-IgG bound to the coated viral antigen was detected by peroxidase conjugated anti-human IgG. In this study, the sensitivity and specificity of developed ELISA was compared with those of commercial ELISA kit and immunofluorescence assay. With these technic 92 sera of healthy pregnant women were tested for the presence of HSV antibodies. The results showed that the sensitivity and specificity of the developed ELISA were 92.5% and 100%, respectively when compared with those of commercial ELISA kit and 100% and 93.75%, respectively when compared with those of immunofluorescence assay. This local-HSV antigen did not show cross reaction with commercial antibody to varicella zoster virus and cytomegalovirus. The prepared antigen was stable upon storage for at least 6 months. The results indicated that the developed ELISA was easy, rapid, sensitive, and specific and therefore, suitable for routine assay of HSV antibody.