Detection of human immunodeficiency virus and other retroviruses in cell culture supernatants by a reverse transcriptase microassay.
A micromethod for the detection of human immunodeficiency virus (HIV) and other retroviruses in cell culture supernatants is described which applies a DEAE ion exchanger for recovery of polynucleotides synthesized in vitro by the retroviral reverse transcriptase. Cell culture, sample preparation, and test performance including the washing step are adapted to microtitre plates. Compared to the standard method this technique produced less non-specific reactions, resulting in a more than 3-fold higher sensitivity, a higher reproducibility due to lower intrarun variations and allowed an increase in the daily sample accomplishment per person 3- to 4-fold at lower costs per sample.
D003636:DEAE-Cellulose; D006678:HIV / Q000201:enzymology* / Q000302:isolation & purification; D006801:Humans; D007474:Ion Exchange; D012194:RNA-Directed DNA Polymerase / Q000032:analysis*; D012190:Retroviridae / Q000201:enzymology* / Q000302:isolation & purification
D003636:DEAE-Cellulose; D012194:RNA-Directed DNA Polymerase