Ultra sensitive method for HIV p antigen detection

Abstract The ultimate goal of the proposed project is to develop a commercial illumination device detection system kit and original ELISA diagnostic kits for detection of HIV p antigen in human serum or plasma by a photochemical amplification method PAM developed within Phases I and II The methods for identifying HIV infection include HIV antibody tests viral isolation through viral culture nucleic acid tests to detect viral RNA tests to detect viral p antigen and tests to detect the viral enzyme reverse transcriptase These methods are highly specific and a positive result almost always confirms infection However each has limitations and their use must be tailored to proper testing situations Tests for viral nucleic acid have been introduced but require sophisticated technology dedicated and well trained personnel and are very expensive Tests to detect p antigen are simpler to perform than nucleic acid tests and offer alternatives for monitoring HIV infection The p antigen test also can be of value in blood screening for identification of acute infection to assist in the diagnosis of infection in the newborn and in detecting antigen in supernatants from cultures that have been inoculated with cells from a patient suspected of being infected At present p antigen tests are relatively insensitive being able to detect only pg ml This quantity of antigen may not be present in the serum of infected individuals even when the virus is actively replicating Therefore development and the use of more sensitive p antigen tests has a great relevance It is in this context that in Phases I and II we developed highly sensitive p antigen assays that are less expensive than RNA viral load tests and appropriate for use both in developed and developing countries in which the limitations of infrastructure and laboratory capability prohibit nucleic acid testing The specific aims of Phase IIB Competing Renewal are To design and manufacture a pre production PAM illumination device To design and develop a PAM detection system kit HIV p ELISA kit and fourth generation ELISA kit for HIV detection as ready to use ELISA PAM diagnostic kits To perform pre clinical studies in accordance with FDA Regulatory pathway using over five hundred clinical samples Narrative The development of the proposed method will allow using HIV p antigen tests for monitoring HIV infection blood screening identification of acute infection to assist in the diagnosis of infection in the newborn and in detecting antigens in supernatants from cultures The performance of the developed tests will be improved significantly as compared with that for conventional assays Successful completion of these studies will be beneficial for public health and make available all of the technology needed for a substantial business opportunity to license the technology and manufacture commercial products