راه‌اندازی و به‌کارگیری روش الیزا به‌منظور تشخیص حضور آنتی‌ژن p24 در خون افراد آلوده به HIV

Introduction: Human immunodeficiency virus is the etiologic agent of immune system deficiency and the infected patients are susceptible to opportunistic infections and AIDS. Transmission of infection during blood transfusion and diagnosis of infection in hospitals and public health settings have steadily been a worldwide concern. Different techniques are now available for the diagnosis of HIV infection. ELISA detection system is used in the diagnosis of antibodies. In order to shorten the window period and detect the infection in newborn, a capture ELISA assay was developed for the diagnosis of HIV capsid or p24 antigenemia in the infected patients. Objective: The aim of this study was to establish and develop of a capture ELISA assay for diagnosis of HIV capsid antigen in serum samples of HIV positive patients. Materials & Methods: In this study polyclonal monospecific antibody against recombinant HIV p24 antigen was developed in rabbits. Then, it was purified by IgG, and p24 specific affinity chromatography. Purified antibody was confirmed and conjugated with conjugated peroxidase “HRP”. Thus, the capture ELISA was developed. To evaluate the sensitivity and the specificity of the technique, HIV positive and negative serum samples were checked and compared to the standard assay. Results: The results indicated that the developed assay could be useful for the detection and confirmation of HIV infection during the window period. The specificity of the assay was %100 and its sensitivity was %96.87. After comparison of findings among subtypes A, B and C of HIV, it was found that there was no statistically significant variation. Conclusion: The developed assay showed high sensitivity and specificity for detection of viral antigen. In addition, it is not time consuming and specific antibody production in animal model is relatively easy. In comparison to other assays, this method is easy to perform, rapid, accurate and inexpensive. The assay could be readily used for the diagnosis of HIV infection in newborns, since the maternal antibodies could pass through the placenta. Moreover, it could to shorten the window period.