Abstract
Acquired immune deficiency syndrome (AIDS) remains a major public health problem in Thailand. Treatment of HIV-infected patient using highly active retroviral therapy (HAART) can suppress viral replication and increases maintain the level of CD4+ lymphocyte cells. Adherence to HAART can improve health and prolong life of HIV-infected patients. However, non-adherence to antiretroviral drugs leads to develop of HIV drug-resistance strains. Therefore, the high costs of new generation antiretroviral drugs are needed. In this study, we aimed to develop a high throughput assay for examining the enzymatic activity of HIV-1 protease (PR) and integrase (IN). These assays will be applicable for assessing the level of HIV-1 protease inhibitors (PIs) and integrase inhibitors in HIV-infected patients who recived antiretroviral treatment. The novel immunochromatographic (IC) strip test for determining of HIV-PR activity was developed by combining proteolysis activity of HIV-PR and an immunochromatographic reaction. This IC strip could adaped to evaluate the biological activity of PI and also semi-quantify the level of PI in HIV-1 infected individual. Furthermore, we established a non-radioisotopic testing based on real-time polymerase chain reaction (PCR) assay to determine HIV-1 integrase activity. This assay has been shown to be effective and inexpensive for a high-throughput screening of novel IN inhibitors. The novel development of enzymatic activity assay of HIV-1 protease and integrase using immunochromatographic strip test and real-time PCR assay could provide as a high-throuput screening platform for discovering new synthetic antiretroviral agents or natural compounds from Thai medicinal plants by Thai reseacher. This study might be able to reduce the importion of new generation antiretroviral drugs from other countires.
