A highly sensitive chemiluminescent reverse transcriptase assay for human immunodeficiency virus.
A simple and highly sensitive reverse transcriptase (RT) assay was developed by combining a previously reported non-radioisotopic RT assay with the use of a template-primer-immobilized microplate, an enzyme capture protocol, product digestion and a chemiluminescent substrate. The assay was able to detect directly the RT activity in serum samples, plasma and cell culture medium without the need for concentration and extraction of the enzyme. The assay was able to detect RT activity equivalent to 100 virions/ml of HIV-1. These results suggest that this highly sensitive chemiluminescent RT assay can be used not only for virological investigation but also for routine screening of biopharmaceuticals.
D002478:Cells, Cultured; D003124:Colorimetry; D003850:Deoxyribonuclease I / Q000378:metabolism; D015658:HIV Infections / Q000821:virology; D054303:HIV Reverse Transcriptase / Q000097:blood*; D015497:HIV-1 / Q000201:enzymology* / Q000502:physiology; D006801:Humans; D008163:Luminescent Measurements; D012367:RNA, Viral / Q000032:analysis; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012680:Sensitivity and Specificity; D013601:T-Lymphocytes / Q000821:virology
D012367:RNA, Viral; D054303:HIV Reverse Transcriptase; D003850:Deoxyribonuclease I