Improved HIV-1 Viral Load Determination Based on Reverse Transcriptase Activity Recovered From Human Plasma

摘要: Reverse transcriptase (RT) is a crucial enzyme for retrovirus replication, and its presence in the virion indispensable infectivity. This thesis illustrates use of RT activity assays as tools quantitation characterization different retroviruses, particularly HIV. A non radioactive assay, using microtiter plates, Moloney murine leukemia virus (MMuLV) was developed. Assay conditions MMuLV HIV-1 RT, together with isozyme specific blocking antibodies, were shown useful discrimination between RTs from genera. assay found to quantitate subtypes more equally efficient than p24 antigen did.Viral load (VL), amount HIV particles blood, an important marker clinical status infected person. method VL determination based on (ExaVir Load) After plasma pretreatment, inactivate cellular DNA polymerases, virions patient immobilized gel, which washed remove disturbing factors. The lysed detergent containing buffer lysate eluted. Finally, determined correlate strongly by RNA according PCR standard (Roche Amplicor 1.5). second version able measure down approximately 400 copies/ml. usefulness procedure susceptibility towards anti-HIV drugs demonstrated, results agreement genotypic data. Due technical simplicity, ability detect broad range subtypes, ExaVir Load drug application are interesting use, but not only resource limited settings. concept also potentially research purposes, e.g. combination conditions.